Breath testing devices and methods

Abstract

The present invention provides a universal breath testing platform and methods of testing for diseases of the gastrointestinal tract, the liver, the kidneys, and the lungs, along with testing for cancer, infections, and metabolic diseases. In certain embodiments, the present invention provides a breath testing platform to predict and monitor athletic performance and health parameters in subjects involved in exercise, gymnastics and sports.

Description

[0001] BREATH TESTING DEVICES AND METHODS

[0002] RELATED APPLICATIONS

[0003] 1. This application is a provisional patent application.

[0004] FIELD OF THE INVENTION

[0005] 2. The present application relates generally to a breath testing device and to methods of detecting multiple breath gases and Volatile Organic Compounds (VOC) present in breath, to determine the presence of diseases, athletic performance and other conditions related to the human microbiome. The present application also relates to utilization of generative and predictive artificial intelligence methods, including but not limited to Machine Learning algorithms, to derive results about diseases present in a person’s body, the performance of a person’s body before, during and after physical exercise. In some instances, the disease can include, but is not limited to, II. pylori infection, Celiac Disease and Non-Celiac Gluten Sensitivity, Metabolic dysfunction- associated steatotic liver disease (MASLD) and Metabolic dysfunction-associated steatohepatitis (MAHD), Irritable Bowel Syndrome, Small Intestinal Bacterial Overgrowth, Lactose and Fructose Intolerance, Inflammatory Bowel Disease (IBD), Pulmonary disease, Intestinal infections and Cancer. In some instances, the physical exercise can include sports including by not limited to basketball, baseball, soccer, running, swimming, tennis walking and others.

[0006] BACKGROUND OF THE INVENTION

[0007] 3. Exhaled breath contains many gases and thousands of Volatile Organic Compounds (VOCs). Gases and VOCs in breath can be detected with the use of technologies like gas chromatography-mass spectroscopy (GC-MS), selected ion flow tube mass spectrometry (SIFT- MS), FTIR spectroscopy, ion mobility mass spectrometry, field asymmetric ion mobility spectroscopy, semiconductor chips, carbon nanotubes, metal oxides, doped and non-doped polymers and other types of conductive materials sensitive to various gases. Types of conductive materials are chemical sensitive field effect transistors or floating gate field effect transistors or any other field effect transistors. Other types of gas sensitive sensors are optical, electrochemical, thermochemical and surface acoustic wave (SAW) thin films deposited on conducting material (e.g., gold, platinum, palladium or other metals). Other types of sensors do not provide for conducting material. 4. Gases in exhaled breath are mainly, oxygen (O2) 16%, carbon dioxide (CO2) 4%, nitrogen (N2) 75% and water vapor (5%-6%). In addition, the exhaled breath contains small amounts of argon, hydrogen (H2), ammonia (NH3), acetone, methanol, ethanol and methane (CH4) which are some of the most commonly encountered organic volatile compounds. Other VOCs in exhaled breath are acetaldehyde, 2-propanol, acetonitrile, acrylonitrile, benzene, isoprene, pentane, methylexane, ethane, hydrogen sulfide, triethyl amine, trimethylamine, carbon disulfide, dimethyl sulfide, 1 -Heptene, 1 -Octene, 1 -Nonene, 1 -Decene, octane, nonene, dodecane, cyclohexane, 2- butane, indole, ester, carbon disulfide, pentane, nitric oxide (NO), ethane and propane. Breath VOCs are metabolic byproducts of physiologic or pathophysiologic processes taking place within the digestive tract, small and large intestine, in healthy or in diseased individuals.

[0008] 5. Metabolomics is the field which deals with the byproducts or metabolites of either physiologic or pathophysiologic processes within the human body. Such byproducts are, among others, the volatile organic compounds (VOCs) derived from breakdown of ingested substances by bacteria of the colon in healthy individuals and in individuals with diseases of the gastrointestinal tract, of the kidneys, of the lung, of metabolism, of the liver, and in individuals with metabolic syndrome, obesity, cancer and infections. Analysis of VOCs and their patterns reflects the composition of microbiota and are considered to be markers of these diseases. Detection of VOCs is currently done using technologies such as gas chromatography-mass spectroscopy (GC-MS), selective ion flow tube mass spectrometry (SIFT-MS), field asymmetric ion mobility spectrometry (FAIMS) and electronic noses (e-Nose).

[0009] 6. Technologies like GC-MS, SIFT-MS, and FAIMS are efficacious in detecting VOCs especially derived from colon microbiota. However, the instruments are expensive, large and mostly stationary. The e-Nose technology, a pattern recognition technology, uses various sensors which are less gas-selective and unstable. None of these technologies use individual sensors which detect each VOC in exhaled breath, and none of these instruments are small, hand-held, inexpensive and available for use over-the-counter (OTC) by exhaling in the instrument directly through an opening of the instrument or a through a mouthpiece. Metal oxide sensors either on their own or coupled with GC-MS are not optimal because the majority of them do not operate at room temperature and most of them need to be heated to about 450 degrees C prior to use. 7. Celiac disease is an autoimmune enteropathy precipitated in susceptible individuals by the ingestion of gluten, specifically its immunotoxic component gliadin. Celiac disease enteropathy resolves with complete and lifelong gluten-free diet. Non-adherence or poor adherence to gluten- free diet may lead to small intestinal lymphoma, adenocarcinoma or other immunological diseases, such as diabetes, thyroiditis, hepatitis, etc. Individuals susceptible to celiac disease express autoantibodies such as antiendomysial and anti-tissue transglutaminase. In addition to those with confirmed Celiac disease, there are patients with subclinical or asymptomatic Celiac disease and family members of patients with Celiac Disease who have abnormal small intestinal histology without symptoms and who remain undiagnosed. The undiagnosed patients and those with delayed diagnosis remain on gluten-containing diet and risk serious damage to their intestinal mucosa and life-threatening consequences like cancer.

[0010] 8. The prevalence of Celiac disease has been increasing in parts of the world, where consumption of gluten is common. Currently in the US, the prevalence of Celiac disease is 1%, much higher than its 0.02% prevalence in the early 1990s. There is also increasing prevalence of Non-Celiac Gluten Sensitivity (NCGS), estimated at 3-6% in the US and Europe. Patients with NCGS have no autoantibodies or intestinal mucosal changes of Celiac disease, but have similar symptoms with Celiac disease patients after ingestion of gluten, which improve on gluten-free diet.

[0011] 9. The current gold standard in the diagnosis of Celiac disease is small bowel biopsies of the small intestinal mucosa which are obtained through upper endoscopy. Patients with Celiac disease, who must follow a strict, lifelong gluten-free diet, undergo follow-up endoscopies with biopsies to assess adherence to gluten-free diet and the health of the small bowel mucosa. Other diagnostic methods exist, including blood tests for antiendomysial and anti -transglutaminase antibodies. Currently there is no screening test for patients with symptomatic Celiac disease or with asymptomatic Celiac disease (subclinical Celiac disease and asymptomatic family members) who do not have the reason to undergo endoscopies. These asymptomatic patients however, are at high risk of developing co-morbidities consistent with those of symptomatic Celiac disease patients and potentially cancer. These asymptomatic patients who are mostly family members of patients with Celiac disease can benefit from certain embodiments of the present invention that offer a non- invasive breath test highly sensitive to the hydrogen in the breath after an eight to twelve (8-12) hour overnight fasting.

[0012] 10. Helicobacter Pylori (H. Pylori), which affects two thirds of the world population, is a highly contagious, gram-negative bacterium which causes chronic gastritis, peptic ulcers and can cause gastric cancer and other malignancies like gastric lymphoma (MALToma). It is associated with extraintestinal diseases like anemia, liver disease, gallbladder disease and pulmonary disease like asthma. One of the properties of H. Pylori is its ability to hydrolyze urea into CO2 and ammonia (NH3) by using its abundant enzyme urease as in the following equation:

[0013] 11. CO(NH2)2 + HOH — urease CO2 + 2NH3.

[0014] 12. The gold standard in diagnosis of H. Pylori is a13C urea breath test (UBT) which measures13CO2in breath after ingestion of13C labeled urea as in the following equation:13CO(NH2)2+ HOH — ureases13CO2 + 2NH3. The current13C UBT requires the use of a relatively expensive kit and very expensive (several thousands of dollars) equipment which must be operated by professionals. There are other known methods to diagnose H. Pylori infection, including: 1) upper endoscopy with biopsies and culture of the tissue; 2) upper endoscopy with biopsies and rapid urease test (CLO test); and 3) serum antibodies to H. Pylori. Positive antibodies indicate infection with H. Pylori. However, the antibodies remain positive even after eradication of the bacterium. As a result, the serum antibodies cannot be used to confirm eradication as the13C UBT (and the other methods) can. None of the existing diagnostic methods are inexpensive and hand-held, combining test and device in one.

[0015] 13. Inflammatory bowel diseases are mainly Crohn’s disease and Ulcerative colitis. There are differentiating features for each of these inflammatory bowel diseases as relating to 1) the part of the intestinal tract which is affected by inflammation in each of them (ulcerative colitis limited to the large intestine and Crohn’s disease potentially affecting the entire digestive tract, but mainly the small intestine); 2) the histological changes in the intestinal mucosa; 3) the diagnosis; 4) the treatment; and 5) the prognosis.

[0016] 14. The diagnosis of inflammatory bowel disease and the differentiation between Crohn’s and ulcerative colitis is established with the use of endoscopies of the small and large intestine with biopsies and histological examination of the intestinal mucosa in addition to radiological studies (especially in Crohn’s disease). After the diagnosis is established, appropriate treatment is instituted. Such treatment is lengthy and requires periodic follow-up with endoscopies and biopsies to assess efficacy of the treatment. It has been observed that changes in the colon-derived metabolome (the distinct type of VOCs and other gases contained in breath) in patients with inflammatory bowel disease (Crohn’s and ulcerative colitis) contribute to the development of a pattern of Volatile Organic Compounds (VOCs) in the breath of these patients. These VOCs are acetaldehyde, 2-propanol, acetonitrile, acrylonitrile, benzene, isoprene, pentane, methylexane, ethane, hydrogen sulfide, triethyl amine, trimethyl amine, carbon disulfide, dimethyl sulfide, 1- Heptene, 1 -Octene, 1 -Nonene and 1 -Decene and others.

[0017] 15. Small intestinal bacterial overgrowth (SIBO) occurs in predisposed individuals and presents with symptoms similar to inflammatory bowel diseases such as abdominal pain, diarrhea, anemia and weight loss. The small bowel normally contains a very small number of microbes as compared to the large bowel, which normally contains several trillion of microbes (estimated at 100 billion per milliliter). SIBO occurs when bacteria, which normally ferment carbohydrates in the large bowel, migrate to the small bowel. As a result, fermentation of carbohydrates takes place in the small bowel, which in turn creates the symptoms of abdominal pain, steatorrhea, and abnormally high production of hydrogen which is absorbed and released through the lungs in the breath.

[0018] 16. Conditions which predispose to the development of SIBO are anatomic abnormalities, abnormal motility of the intestine as in pseudo-obstruction, absence of the migratory motor complexes, autonomic neuropathy as in diabetes, excessive bacterial load as in achlorhydria, fistula and loss of the ileocecal valve and immunological problems like immunodeficiency and malnutrition.

[0019] 17. Diagnosis of SIBO is obtained with invasive or non-invasive tests. Invasive tests include small bowel endoscopy and aspiration of content for culture for aerobic and anaerobic bacteria and motility studies. Non-invasive tests are 72-hour stool collection for fecal fat, serum bile acids and breath tests. The most common breath test is the hydrogen breath test after consumption of lactulose. Lactulose, a non-absorbable sugar, provides the substrate for the colonic bacteria to produce hydrogen which results in increased hydrogen levels in breath. If the large bowel bacteria are present in the small bowel then the hydrogen production after ingestion of lactulose occurs much earlier than usual and an early and late peak of hydrogen production is observed during the breath test. The gold standard hydrogen breath test is performed with Quintron breath analyzer equipment. The individual undergoing breath testing for SIBO provides a baseline fasting breath sample and ingests a certain amount (about 7g / Kg) of lactulose. In cases of SIBO, post-lactulose excretion of hydrogen gives two peaks, one early because large bowel bacteria are in the small bowel, and another peak later when the lactulose reaches the large bowel.

[0020] 18. Irritable bowel syndrome (IBS) is a diagnosis of exclusion and is given to individuals who present with symptoms of abdominal pain, diarrhea or constipation, bloating and flatulence. IBS is a functional disorder of the digestive tract which, despite symptoms similar to TBD, has no organic etiology and no histologic abnormality of the digestive tract has been associated with it. Intestinal motility abnormalities and food intolerance have been observed in certain individuals with IBS which can present with either diarrhea or constipation. Diagnosis of IBS is made after all organic causes for IBD, SIBO, infections and Celiac Disease have been ruled out. It is reported that analysis of fecal Volatile Organic Metabolites has a pattern consistent with IBS with diarrhea (IBS-D) which differentiates IBS-D from IBD and healthy individuals with a 94-96% sensitivity and 80-82% specificity. IBS-D fecal volatile organic compounds contain esters of short chain fatty acids such as cyclohexane carboxylic acids, butyrate, acetate and propionate. This observation can lead to developing the colon-derived breath metabolome as markers for IBS especially with diarrhea.

[0021] 19. Lactose intolerance is intolerance to lactose, the carbohydrate contained in milk. Lactose intolerance presents with symptoms similar to Irritable Bowel Syndrome and other malabsorption problems. The symptoms are abdominal pain, diarrhea and flatulence and occur after ingestion of milk or milk-containing products. Lactose is a disaccharide which is broken down to the monosaccharides glucose and galactose with the use of the enzyme lactase which is an enzyme present at the intestinal brush border. Glucose and galactose are easily absorbed and usually cause no symptoms. In individuals in whom the enzyme lactase is absent or severely limited, as in genetic conditions or post infections involving the digestive tract like, viral enteritis or parasitic or microbial infections, lactose is not metabolized and as a result it causes symptoms similar to carbohydrate malabsorption, diarrhea, abdominal pain, bloating and flatulence. Fructose intolerance is the inability to tolerate intake of fructose, the carbohydrate found in fruits. As a result, individuals with such inabilities who ingest fruits experience similar symptoms as the individuals with lactose intolerance. It is estimated that the adult human intestine can tolerate up to 25g of fructose without symptoms. Ingesting more than 25 grams of fructose can cause diarrhea, flatulence and abdominal pain. Any amount of fructose can cause symptoms in individuals who have the genetic form of fructose intolerance due to genetic deficiency of the enzyme Aldolase B.

[0022] 20. Liver diseases are divided in two categories; genetic and inherited liver diseases and acquired liver diseases. The genetic and inherited liver diseases include disorders of carbohydrate metabolism, disorders of amino acid metabolism, abnormalities in mitochondrial fatty acid P- oxidation, al-antitrypsin deficiency, disorders of the bile acid synthesis and metabolism, Wilson disease and others. The acquired liver diseases are mainly viral infections like hepatitis A, B, C and others which can lead to chronic hepatitis, cirrhosis and end-stage liver disease, Hepatocellular carcinoma (HCC) and other liver malignancies and diseases of the liver due to obesity and alcohol use, such as metabolic dysfunction associated steatotic liver disease (MASLD) aka NAFLD and metabolic dysfunction-associated steatohepatitis (MASH) aka NASH and alcoholic liver disease (ALD) and metabolic abnormalities such as diabetes and others. Autoimmune hepatitis, which can lead to chronic hepatitis and cirrhosis, is an inflammatory hepatitis which is secondary to autoimmune diseases affecting more than one body organ. HCC can be the result of viral hepatitis and of steatohepatitis which is due to deposits of fat in liver tissue. Steatohepatitis can be secondary to obesity, diabetes and alcoholic hepatitis. If untreated, steatohepatitis can lead to cirrhosis and end stage liver disease. Changes in metabolome have been observed in genetic liver diseases as well as in acquired liver disease and in end stage liver disease. Hyperammonemia induced encephalopathy is one of the symptoms of liver failure.

[0023] 21. Renal diseases are diseases of the kidneys and are genetic-idiopathic and acquired. Genetic diseases of the kidneys include several types of glomerulonephritis such as IgA nephropathy, Alport syndrome, Goodpasture disease, membranous glomerulopathy, lupus-induced glomerulonephritis and others and anatomic abnormalities such as polycystic disease of the kidneys and UPJ obstruction. Acquired kidney diseases are post-infection glomerulonephritis such as acute post-streptococcal glomerulonephritis, hemolytic uremic syndrome (HUS), drug-induced glomerulonephritis, post-traumatic renal disease and others. Symptoms of renal disease are hematuria, proteinuria, edema and poor growth in children. When renal disease becomes chronic and progresses to renal failure (kidney failure), hemodialysis and eventual kidney transplant are essential. Hyperammonemia is a key feature of end stage renal failure due to the inability of kidneys to excrete the accumulating ammonia.

[0024] 22. Pulmonary disease. More than 3,500 components (gases) make up the human breath and the majority are volatile organic compounds (VOCs) in very small amounts. In addition to nitric oxide (NO), which has become a biomarker for respiratory inflammation and asthma, a pattern of VOCs has emerged as prevalent within the population with allergic and inflammatory component asthma. These compounds are nonane, 2,2,4,6,6-pentamethylheptane, decane, 3,6- dimethyldecane, dodecane, and tetradecane. The breath metabolome provides a characteristic of chronic obstructive pulmonary disease (COPD).

[0025] 23. Metabolic diseases like diabetes and obesity affect the metabolome. Increased levels of acetone in breath of individuals with diabetes, is a finding consistent with undiagnosed diabetes or poorly managed diabetes in individuals who are already diagnosed. The concentration of breath acetone has been found to correlate with the P-hydroxybutyrate concentration of venous blood in fasting obese patients. Studies have identified eight specific metabolites such as isopropanol and 2,3,4-trimethylhexane, 2,6,8-trimethyldecane, tridecane and undecane as a more specific pattern for the presence of type 2 diabetes with a high sensitivity and specificity. Increase in VOCs has been described in patients with various types of cancer such as colon, lung, breast and others. Infections involving the gastrointestinal tract such as bacterial infections (salmonella, shigella), parasitic infections and Clostridium Difficile alter the colonic bacteria, and as a result, alter the corresponding breath metabolome.

[0026] 24. Helicobacter pylori ^H. pylori”) is a gram-negative bacterium found in the digestive tract that affects about two-thirds of the world’s population. Most people contract H. pylori infection during childhood and may never have any signs or symptoms of the infection. However, when signs or symptoms do occur with H. pylori infection, they may include a burning pain in the abdomen and chest, nausea, vomiting, frequent burping, bloating and weight loss. H. pylori causes gastritis and the majority of peptic ulcers, and is associated with other intestinal diseases like inflammatory bowel diseases, hepatobiliary and pancreatic diseases. H. Pylori is also associated with extraintestinal diseases, some of which are esophageal strictures, idiopathic thrombocytopenic purpura, iron deficiency anemia, renal diseases, and cardiovascular diseases like ischemic heart disease and atherosclerosis.

[0027] 25. In addition, H. Pylori was declared a Class I carcinogen for humans in 1994 by the International Association for Research on Cancer (IARC) and has been strongly associated with the development of gastric cancer and with gastric MALToma, a type of lymphoma. H. Pylori has also been associated with other cancers like colon cancer and pancreatic cancer.

[0028] 26. Current diagnostic methods for detecting H. Pylori infection have several drawbacks. Some existing diagnostic methods include blood tests, endoscopy with biopsies and bacterial culture. Such methods are high risk, expensive, uncomfortable. In addition, the results of such tests are not available until several days after the test has been performed. Furthermore, these invasive methods utilize large and expensive equipment operated by highly-trained personnel; require the use of either anesthesia, sedation, or both; and require the use of pathology and microbiology laboratories to render a diagnosis.

[0029] 27. Currently available non-invasive methods for the detection of H. Pylori include the13C labeled urea breath test (13C-UBT); detecting H. pylori antigens in stool; a stool test for H. Pylori DNA. These known non-invasive methods are costly, not available for self-testing, and the testing results are unavailable until several hours or days after the test has been performed. Currently, these non-invasive methods utilize centralized laboratory equipment for analysis of the breath sample for13CO2 and to examine stool for the H. Pylori antigen and / or for H. Pylori DNA. All of these methods require expensive equipment and qualified personnel to carry out the particular testing method. Some of these methods are messy and less sensitive and specific for the presence of bacterium.

[0030] 28.13C-Urea breath tests are also known and have been used to detect H. pylori infection. In such known types of urea breath tests, the presence of H. Pylori is based on the measurement of the ratio of13CO2:12CO2 in the breath of a subject after ingestion of a urea substrate labeled with13C. The subject exhales into a bag, which is then attached to large equipment (e.g., a spectrometer) for analysis of the breath sample. If the13CO2:12CO2 ratio is above a certain value, the subject is considered positive for H. pylori. However, these known types of urea breath tests have several drawbacks. First, they use a meal which contains urea urea labeled with13C, which, though it is a naturally occurring isotope of carbon, is expensive in comparison to the also naturally occurring12C which is contained in the unlabeled urea. In addition, known13C-Urea breath tests require expensive instruments (e.g., infrared spectrometers, GC-mass spectroscopy) to analyze13CC>2 and12CO2levels.

[0031] 29. As set forth in the present disclosure, it would be desirable to provide a breath test method that measures elevated levels of both ammonia and CO2(measured as12CO2or13CO2; both12CO2and13CO2; or as a ratio of13CO2:12CO2) in the breath of a subject before and after the subject ingests a urea substrate. It would also be desirable to provide such a breath test that can utilize a urea substrate regardless of whether the urea is labeled or unlabeled. In addition, it would be desirable to provide a reliable point-of-care diagnostic test that can be self-administered using an autonomous hand-held device to determine the presence of H. pylori. Still further, it would be desirable to provide an improved breath test that is less costly and more convenient than existing breath tests.

[0032] 30. Breath testing for the detection and measurement of biomarkers for disease. The breath metabolome, which is a constellation of gases found in the breath, can be used as biomarkers for several diseases of the digestive tract, liver, pancreas, kidneys, lungs as well as cancer and infections.

[0033] 31. One breath biomarker is fasting breath hydrogen (FBH), a diagnostic breath biomarker for Celiac disease. Fasting breath hydrogen (FBH) is significantly higher in patients with Celiac disease than in patients with other malabsorption syndromes and remains elevated until the patient remains on strict gluten-free diet for at least 6 months. FBH can be used for monitoring of patients with Celiac disease on strict gluten-free diet because, it has been shown, that the level of FBH correlates with the histologic condition of the small bowel mucosa. The higher the FBH the more abnormal the histology of the small bowel, and the reverse, the lower the FBH the less abnormal the histology of the small bowel. When the FBH is within normal limits, the histology of the small bowel is normal.

[0034] 32. Another breath biomarker is fasting breath ammonia (FBA). MASH is part of the spectrum of metabolic dysfunction-associated steatotic liver disease (MASLD), which has become the most common liver disease in the world due to worldwide increasing rates of obesity and metabolic syndrome. MASH develops when lipotoxic lipids, accumulated due to fatty infiltration of the liver, cause significant hepatocellular injury. Lipotoxic lipids (diacylglycerols, ceramides and others) mediate an array of intracellular processes (endoplasmic reticulum stress, mitochondrial dysfunction, inflammation and apoptosis) to produce the histologic phenotype of MASH. These intracellular processes are the stimuli for fibrogenesis and possibly hepatocellular carcinoma (HCC).

[0035] 33. An estimated 2-5% of the US population or 20% of the population with obesity-induced MASLD will advance to MASH and cirrhosis without clearly established predictability criteria and MASH may soon become the leading indication for liver transplantation. The cumulative annual risk for developing HCC in patients with MASH-fibrosis, if untreated, ranges from 2.4% to 12.8% and the projected annual economic impact of MASLD-MASH is estimated at $103 billion in the US. There is a dire need to identify and treat MASH before the development of cirrhosis.

[0036] 34. Liver biopsy, an invasive procedure, is the gold standard in diagnosis and staging of MASH fibrosis along with elevated serum alanine transferase (ALT) >50 IU / L and imaging (ultrasonography-MRI) studies. Biomarkers are expected to improve the ability to stratify disease severity in MASLD and to identify additional pathways to target for treatment before it advances to MASH fibrosis and cirrhosis. Rigorous review of the literature revealed that several non- invasive biomarkers and panels of biomarkers with blood tests that reflect underlying disease pathways in MASH are being developed. Biomarkers such as Caspase-generated CK-18 fragment (CK-18), fibroblast growth factor 21(FGF21), insulin-like growth factor 2 (IGF-2) as well as epidermal growth factor receptor (EGFR) have moderate diagnostic and prognostic accuracy. Biomarker panels such as AST: ALT ratio, MASLD fibrosis score, BARD score (BMI, AST: ALT ratio, Diabetes mellitus) are less accurate than specific fibrosis markers (FibroTest, FibroMeter). These specific fibrosis markers accurately predict the fibrosis stage, only, after fibrosis has long been developed. Hyaluronic acid (HA), a major component of extracellular matrix, is detected during advanced stages of fibrosis.

[0037] 35. Imaging biomarkers such as FibroScan and point shear wave elastography (psWE) have moderate to high accuracy of diagnosing advanced fibrosis or cirrhosis but not early stages of fibrosis. Magnetic Resonance Elastography (MRE) has higher success rate and accuracy than ultrasound-based technologies but is limited by cost and availability. Genetic and genomic markers for assessment of disease susceptibility and disease severity are being developed but with limitations in accuracy and reproducibility with regards to prediction of NAFLD and NASH and its severity. Metabolomic studies are underway and studies of the microbiome profile produced an algorithm that could predict advanced fibrosis suggesting that a test based on it would be a useful biomarker. However, such marker would not predict early fibrogenesis. So far, despite promising clinical studies, an accurate, reproducible non-invasive method to predict early NASH has not been identified.

[0038] 36. There is a need for accurate, non-invasive, reproducible, and specific biomarker with high predictive value of NASH fibrosis at the earliest possible stage of fibrogenesis.

[0039] 37. Ammonia in MASH: None of the existing biomarkers is a breath test, the simplest method of testing for a potential biomarker. Recent, compelling evidence in animal studies and in humans shows that hepatocellular injury causes structural changes (gene expression, activity) in the urea cycle enzymes ornithine transcarbamylase (OTC) and carbamoyl phosphate synthetase (PS), thus impairing ammonia’s conversion to urea resulting in hyperammonemia. Studies have shown a direct link between increased ammonia concentration and the development of MASH fibrosis. Ammonia at concentrations of 50-300 pM / L activates the usually quiescent hepatic stellate cells (HSC) which then become highly proliferative and synthesize a fibrotic matrix rich in type I collagen becoming key for the development of fibrosis, portal hypertension and HCC. Hyperammonemia due to hepatocellular injury is among the earliest stimuli for the development of fibrosis in MASH.

[0040] 38. Breath ammonia as biomarker for MASH: The toxicity of hyperammonemia makes monitoring of ammonia a priority. However, measurement of blood ammonia is considered nonreliable, as per AASLD, due to inaccuracies associated with blood drawing methods (arterial blood is a preferable but painful procedure and requires professional training- Venus blood often gives inaccurate results) and errors with the transport of the specimen to the lab. Measurement of breath ammonia, a simpler and more accurate procedure than blood ammonia, can replace blood ammonia especially since ammonia is in equilibrium in the lungs and studies have shown that blood ammonia correlates with breath ammonia.

[0041] 39. Clostridium Difficile (C. Diff.) infection. C. Diff infection of the intestine, which presents as abdominal pain, diarrhea and weight loss, is the result of antibiotic use and due to compromise of the colonic mucosa in individuals with diseases affecting the microbiome. As a result, the breath metabolome of these affected individuals differs from the ones that are not affected. Studies have shown that a pattern of VOCs is observed in patients with C. Diff infection. Such VOCs are propan- l-ol, 3 -methylbutanal, ethyl propionate, hexanoic acid, 4-methyl phenol, dodecane, and indole. Other studies, addressing the different ribotypes of the bacterium, have identified methanol, p-cresol, dimethylamine and a range sulfur compounds (ethylene sulfide, dimethylsulfide and methyl thioacetate) as VOCs to differentiate such ribotypes.

[0042] 40. Colorectal cancer. One of the most common cancers in the United States is colorectal cancer (CRC), which has begun affecting young as well as older individuals, is largely preventable by periodic screening colonoscopy for removal of potentially cancerous polyps. Studies have identified breath gases and VOCs associated with CRC. These gases, mainly 1 -iodononane and benzene, although others like Nonanal, 4-Methyl-2-pentanone, Decanal, 2-Methylbutane, 1,2- Pentadiene, 2-Methylpentane, 3 -Methylpentane, Methylcyclopentane, Cyclohexane, Methyl cyclohexane, 1,3 -Dimethylbenzene, 4 -Methyl octane, 1,4-Dimethylbenzene, 4- methylundecane, trimethyldecane, have been associated with CRC. An autonomous, hand-held breathalyzer device and a breath test to screen for colorectal cancer based on VOC breath biomarkers will save lives and will lower health care cost.

[0043] 41. Athletics-gymnastics-sports. Exercise, gymnastics and sports in general create the metabolic conditions within the human body, which produce changes of the concentrations of breath gases and VOCs. During exercise oxygen is utilized by the body by large amount and CO2 is also produced at higher concentrations than normally present in breath at rest. Additionally, acetone and isoprene have been reported to undergo changes post exercise. Isoprene has been found to decrease significantly after 10 minutes of exercise and to return to the baseline at 60 minutes.

[0044] 42. Predicting performance by athletes is desirable and has been based on psychological and actual performance measurements on an individual athlete basis. Such prediction is desirable by leadership of sports teams and has been applied to date by coaches. Breath gases and VOCs have not been used routinely as predictive factors for athletic performance. Breath testing, utilizing mass spectroscopy equipment, has demonstrated the changes in breath gases before, during and after exercise. Such measurements, however, have not been used to predict performance by athletes. Software with embedded Machine Learning algorithms, can, based on selected parameters, assist in identifying and accurately classifying athletic performance based on data, which include performance parameters, anthropometries and biological parameters such as exhaled breath gases and others.

[0045] 43. Medical wearables is a $36 billion worldwide market projected to increase at an annual growth rate of 27.5% from 2024 through 2029 (reports). Autonomous wearable devices, e.g., the apple watch, offer the opportunity to users and healthcare providers to establish healthcare channels for easier monitoring and communication. One segment of the wearables is the exercise / athletics segment, another is breath diagnostic. Autonomous breath testing wearables will provide breath testing using miniaturized, light-weight, fitting equipment can be placed generally on the face to access the breath. Other parts of the body can accommodate wearable breath testing device, e.g., the wrist. Miniaturized electronic boards and miniaturized gas-selective sensors will fit in small-size appropriate for wearing on parts of the human body, e.g., the face and the wrist.

[0046] SUMMARY OF THE INVENTION

[0047] 44. Certain embodiments of the present disclosure provide an autonomous universal, electrochemical multi-analyte breathalyzer device for the detection of gases and VOCs in human breath of individuals with gastrointestinal, renal, respiratory, metabolic, inflammatory diseases, cancer, and infections. The universal electrochemical device can comprise a main body which houses one or multiple electrochemical sensors, software with machine learning algorithms embedded, a processor, the electronics to support the operation of the device, a power source, a USB port, and Bluetooth technology to transfer data wirelessly to another device or computer or the Cloud. The universal electrochemical device, which can have a touchscreen display for input by the user and for output of the results, accepts the user’s breath through either an opening of the body of the device (which acts as mouthpiece) or through a removable and replaceable mouthpiece.

[0048] 45. Certain embodiments provide an autonomous, hand-held breath analyzer that includes an input, a first sensor, a second sensor, software and a processor, and an electrical circuit. The input receives the breath sample. The first sensor contacts the breath sample and includes polyaniline and a conductive material. The polyaniline contacts the conductive material and is doped with a first dopant that increases pH sensitivity of the polyaniline. The polyaniline has a resistivity that increases in response to increased concentration of ammonia. The second sensor also contacts the breath sample. The second sensor includes polyaniline and a conductive material. The polyaniline contacts the conductive material and is doped with a second dopant that increases pH sensitivity of the polyaniline. The doped polyaniline has a resistivity that increases in response to increased concentration of carbon dioxide. The electrical circuit operably connects the first and second sensors to the processor. The processor detects resistivity in the electrical circuit and uses the resistivity to calculate the concentration of ammonia and the concentration of carbon dioxide in the breath sample.

[0049] 46. Other embodiments provide an autonomous handheld, portable breath analyzer that includes a removable mouthpiece and a main body. The main body includes a first sensor, a second sensor, software and a processor, and an electrical circuit. The first sensor includes polyaniline, and the polyaniline is doped with a first dopant that increases pH sensitivity of the polyaniline. The polyaniline has a resistivity that increases in response to increased concentration of ammonia gas. The second sensor comprises polypyrrole, and the polypyrrole is doped with a second dopant that increases pH sensitivity of the polypyrrole. The polypyrrole has a resistivity that increases in response to increased concentration of carbon dioxide. The electrical circuit operably connects the first sensor and the second sensor to the processor. The processor detects resistivity of the first sensor and uses the resistivity to calculate a concentration of ammonia, and the processor detects resistivity of the second sensor and uses the resistivity to calculate a concentration of carbon dioxide.

[0050] 47. Still other embodiments provide a breath test method. The breath test method includes the step of providing a portable breath analyzer that includes a removable mouthpiece and a main body. The main body includes a first sensor, a second sensor, software and a processor, and an electrical circuit. The first sensor includes ammonia selective material that has a resistivity that increases in response to increased concentration of ammonia gas. The second sensor comprises carbon dioxide selective material that has a resistivity that increases in response to increased concentration of carbon dioxide gas. The electrical circuit operably connects the first sensor and the second sensor to the processor, and the processor measures resistivity of the first sensor and the second sensor. The method further includes prompting a subject to exhale a baseline breath sample into the removable mouthpiece, and allowing the processor to measure a resistivity of the first sensor that occurs when the baseline breath sample contacts the first sensor. The method also includes prompting a subject to exhale a post-urea-ingestion breath sample into the removable mouthpiece, and allowing the processor to measure a resistivity of the first sensor that occurs when the post-urea-ingestion breath sample contacts the first sensor. Additionally, the breath test method includes the step of comparing the measured resistivity of the baseline breath sample to the measured resistivity of the post-urea breath sample.

[0051] 48. Certain other embodiments provide a method of detecting presence of H. pylori in a digestive tract of a subject. The method includes collecting a fasting baseline breath sample from a subject and determining the amount of ammonia and the amount of carbon dioxide present in the baseline breath sample. The method further includes collecting a post-urea-ingestion breath sample from the subject, and determining a total amount of ammonia and a total amount of carbon dioxide present in the post-urea breath sample. Additionally, the method includes the step of designating a presence of H. pylori in the digestive tract if the amount of ammonia and the amount of carbon dioxide present in the post-urea-ingestion breath sample exceeds the amount of ammonia and the amount of carbon dioxide present in the baseline breath sample by a predetermined amount.

[0052] 49. Certain embodiments provide a breath test method, which includes the step of providing an autonomous, hand-held, portable breath analyzer that includes a removable mouthpiece and a main body. The main body includes a sensor, software and a processor, and an electrical circuit. The electrical circuit operably connects the first sensor and the second sensor to the processor, and the processor measures resistivity of the sensor. The sensor includes ammonia selective material that has a resistivity that increases in response to increased concentration of ammonia gas. Certain embodiments provide a method of detecting presence of Metabolic Dysfunction-Associated steatotic liver disease (MASLD) and / or MASH in the liver of a subject.

[0053] 50. Certain embodiments provide a breath test method. The breath test method includes the step of providing an autonomous, hand-held breath analyzer device which comprises a removable and replaceable mouthpiece and a main body. The main body contains a first sensor and a second sensor, a processor and an electric circuit. The electrical circuit operably connects the first and the second sensor to the processor and the processor measures resistivity of the sensors. The first sensor includes hydrogen selective material that has a resistivity that increases in response to increase of hydrogen concentration. The second sensor includes methane selective material that has a resistivity that increases in response to the increase of methane concentration. Through the removable and replaceable mouthpiece, the exhaled breath comes in contact with the first sensor and with the second sensor. 51. Certain embodiments provide for a breath test method to detect the presence of carbohydrate malabsorption in a subject. The carbohydrate malabsorption can be the result of lactose and / or fructose intolerance of the subject or the result of Small Bowel Bacterial Overgrowth in a subject. The breath test method includes prompting the subject to exhale a baseline breath sample into the breath testing device through the removable and replaceable mouthpiece. The breath test method also includes the subject to ingest carbohydrate substrate and exhale a post-carbohydrate-substrate ingestion breath sample into the breath testing device though the removable and replaceable mouthpiece. The breath test method also includes the calculation of the post-carbohydrate-substrate ingestion concentration of hydrogen and methane and the calculation of the baseline concentration of hydrogen and methane. The breath test method also includes the calculation of the equation post-carbohydrate-substrate ingestion concentration of hydrogen and methane minus the baseline concentration of hydrogen and methane. When the result of the equation is greater than a predetermined value, the breath test method detects positive result. When the result of the equation is less than a predetermined value, the breath test method detects negative result.

[0054] 52. Certain embodiments provide for a breath test method. The breath test method includes the step of providing an autonomous, hand-held breath analyzer device which comprises a removable and replaceable mouthpiece and a main body. The main body contains a first sensor, a second sensor, a third sensor, a fourth sensor, a fifth sensor and a sixth sensor, software with machine learning algorithms embedded and a processor and an electrical circuit. The electrical circuit operably connects the six sensors to the processor and the processor measures resistivity of the sensors. The first sensor includes hydrogen selective material that has a resistivity that increases in response to increase of hydrogen concentration. The second sensor includes ammonia selective material that has a resistivity that increases in response to the increase of ammonia concentration. The third sensor includes carbon dioxide (CO2) selective material that has a resistivity that increases in response to the increase of CO2 concentration. The fourth sensor includes methane selective material that has a resistivity that increases in response to the increase of methane concentration. The fifth sensor includes acetone selective material that has a resistivity that increases as a result of the increase of acetone concentration. The sixth sensor includes ethanol selective material that has a resistivity that increases as a result of increase of the ethanol concentration. The exhaled breath comes in contact with every one of the six sensors. 53. Certain embodiments provide for a breath test method to prompt a subject to exhale a fasting baseline breath sample into the breath testing device through the removable and replaceable mouthpiece. The breath test method provides for the subject to abstain from food for 8-12 hours and then brush their teeth 15 minutes before taking the breath test. Further the breath test method provides for the subject to exhale a breath sample into the breath testing device through the removable and replaceable mouthpiece. In certain embodiment the breath test method detects the presence of metabolic syndrome in the body of the subject.

[0055] 54. Principal Component Analysis (PCA) is an analytical method, which can be used in medical testing to classify data from clinical studies. PCA can identify subjects with the same diagnosis and cluster them together on the basis of input of components included in the various studies.

[0056] 55. Certain embodiments provide for autonomous, hand-held breath testing device which includes seven or more sensors, software with embedded machine learning algorithms and a processor and electrical circuit. The electrical circuit connects operably each sensor with the processor and the processor detects each sensor’s resistivity and converts resistivity to the concentration of gas for which each sensor is selective. The software provide for increased accuracy and classification of breath testing data. Artificial intelligence and machine learning algorithms are utilized to better interpret the data and the results of breath testing methods. Machine learning algorithms are utilized to examine clustering of data and detect data with precision in way that is not accomplished by utilizing standard statistical methods. Machinelearning algorithmic methods include but are not limited to K-means, DBSCAN or KNN. Certain embodiments include algorithms to improve interpretation of data derived from anthropometric, physical, psychological, clinical and breath testing of a subject.

[0057] 56. Certain embodiments provide for a breath test method. The breath test method includes a step of providing an autonomous, hand-held breath testing device which includes a removable and replaceable mouthpiece and a main body. The main body includes 6 or more sensors, software embedded with machine learning algorithms and a processor, and an electrical circuit. The first sensor includes a nitrogen selective material which has resistivity which increases as a result of the increased concentration of nitrogen. The second sensor includes a CO2 selective material with resistivity which increases in response to increased concentration of CO2. The third sensor includes oxygen selective material that has resistivity which increases in response to increased concentration of oxygen. The fourth sensor includes isoprene selective material that has a resistivity which increases in response to increased concentration of isoprene. The fifth sensor includes acetone selective material that has a resistivity which increases in response to increased concentration of acetone. The sixth sensor includes methanol selective material that has a resistivity which increases in response to increased concentration of methanol. The seventh sensor includes nitric oxide selective material that has a resistivity which increases in response to increased concentration of nitric oxide. In certain embodiments the breath test method provides for predicting the athletic performance of a subject. The method includes collecting a breath sample from the subject before, during and after exercise. The exercise includes but is not limited to swimming, cycling, running, playing tennis, basketball, baseball, football, soccer and other Olympic sports. The breath sample contacts each of the seven sensors and the breath testing device measures concentration of nitrogen, oxygen, CO2, isoprene, acetone, methanol, nitric oxide, in breath, before, during and after exercise.

[0058] 57. Certain embodiments provide for autonomous wearable breath testing device. The wearable device includes one or more than one gas-selective sensors, software and processor and electrical circuit. The electrical circuit operably connects the sensors with the processor. The one or more than one sensors comprise of breath gas selective material that has resistivity that increases as result of increased concentration of the breath gas to which it is selective. In certain embodiments one sensor is selective of two or more breath gases. In other embodiments the autonomous wearable breath testing device collects the data from the user, analyzes the data, stores the data, calculated the results, displays the results on the device and transmits the data and results to smart phone, or computer, or the Cloud.

[0059] 58. In certain embodiments the autonomous wearable breath testing device is a face mask (Fig. 38) manufactured with material to allow breath to flow through and reach the sensors, which are connected to the processor through the electrical circuit. The mask, which can include one or more layers, can be worn over the mouth and the nose, or only over the mouth, or only over the nose.

[0060] 59. In certain embodiments the autonomous wearable breath testing device is worn around a body part, e.g., the wrist or the arm, like a watch (Fig. 39). The device comprises a removable mouthpiece and a main body that includes two parts. One part contains the sensor or sensors, the software and processor and the electrical circuit and the other part contains the mouthpiece opening and the screen. In other embodiments the main body includes one part that contains the sensors, the software and processor and the electrical circuit. In other embodiments the device includes more than two parts.

[0061] BRIEF DESCRIPTION OF THE DRAWINGS

[0062] 60. Figure 1 shows a schematic of a breath analyzer according to an embodiment of the present disclosure.

[0063] 61. Figure 2 shows another schematic of a breath analyzer according to an embodiment of the present disclosure.

[0064] 62. Figure 3 shows a schematic of a breath analyzer according to yet another embodiment of the present disclosure.

[0065] 63. Figure 4 shows an embodiment of an ammonia sensor.

[0066] 64. Figure 5 is a graph showing a change in resistivity of a polyaniline-CSA sensor when the sensor comes into contact with pure gas.

[0067] 65. Figure 6 is a graph showing measurement of13CC>2 concentration in breath samples of multiple subjects, after the subjects ingested a high-protein meal followed by 5 mg / kg of13C- labeled urea.

[0068] 66. Figure 7 shows an embodiment of an electrical schematic for the breath analyzer.

[0069] 67. Figure 8 shows another embodiment of an electrical schematic for the breath analyzer.

[0070] 68. Figure 9 shows a schematic of an embodiment of a breath analyzer.

[0071] 69. Figure 10 shows an embodiment of a breath analyzer with a mouthpiece and a main body in a detached configuration.

[0072] 70. Figure 11 shows an embodiment of a breath analyzer with a mouthpiece and a main body in an attached configuration.

[0073] 71. Figure 12 shows an embodiment of a desiccant assembly. 72. Figure 13 shows an embodiment of a desiccant assembly.

[0074] 73. Figure 14 shows the exterior of a multi-analyte breathalyzer device.

[0075] 74. Figure 15 shows an embodiment of the exterior of a multi -analyte breathalyzer device having an example menu after the menu icon is selected by the user.

[0076] 75. Figure 16 shows an embodiment of an interior of a multi -analyte breathalyzer device having 21 sensors. The device can contain more sensors as needed. This version of the device has an opening which serves as mouthpiece and accepts the breath sample from the user who applies the mouth around the opening. The opening contains several desiccant crystals embedded in the wall at the opening but the mouth does not come in contact with the crystals. The interior of the device contains an electronic circuit and a microprocessor connected to each sensor, along with Bluetooth, a USB port, a battery and a small hole for release of breath as it exits the device.

[0077] 76. Figure 17 shows a display of multi -analyte breathalyzer device with an example of the menu after the icon is selected by the user. The user can tap any of the icons which are shown on the display or the icon which says complete profile to receive measurement of all gases which can be measured by the device. A mouthpiece with desiccant and / or filter embedded in the wall of the mouthpiece can be replaceable.

[0078] 77. Figure 18 shows the display of the multi-analyte breathalyzer device after the user selects one of the icons shown in Figs. 15, 17. The gas or gases to be measured are displayed as are the instructions to the user and proposed follow-up. This version of the device does not contain desiccant / filter inside the mouthpiece. In this version of the device, the desiccant / filter is inside the body of the device in the immediate vicinity of the sensor(s).

[0079] 78. Figure 19 shows one version of the display of the multi-analyte breathalyzer device after the completion of a breath test. Provided are the results in a form of concentrations of detected gas(es) which were selected to be measured by the device according to the selection by the user from the menu. The device also provides the assessment of the results (normal, below or above normal or inconclusive) based on the selection by the user from the menu and based on the demographics input by the user, and provides recommendations for follow-up.

[0080] 79. Figure 20 shows an embodiment of an interior of a single-sensor breath analyzer device. A single sensor is placed inside the device along with electronics and the microprocessor. The device can operate with the use of a battery and can have Bluetooth and a USB port. The device includes an opening for the user to exhale into the device, and desiccant and / or filter are embedded in the wall of the device at the opening.

[0081] 80. Figure 21 shows an interior of one version of the multi -analyte breathalyzer device with two sensors. As an example, one sensor can detect ammonia and the other sensor can detect CO2. This version of the device detects both gases from the same breath sample either one time (one breath sample) or multiple times (multiple breath samples), stores the concentration of gas(es) from each measurement and calculates and stores the differences in values between the various measured concentrations according to a predetermined order.

[0082] 81. Figure 22 shows the interior of one version of the multi -analyte breathalyzer device which contains twenty one (21) sensors. As an example, each sensor can detect each of the following gases and VOCs : The VOCs are acetaldehyde, 2-propanol, acetonitrile, acrylonitrile, benzene, isoprene, pentane, methylexane, ethane, hydrogen sulfide, triethyl amine, trimethyl amine, carbon disulfide, dimethyl sulfide, 1 -Heptene, 1 -Octene, 1 -Nonene and 1 -Decene and others.

[0083] 82. Figure 23 shows the exterior of a multi-sensor, multi-analyte device.

[0084] 83. Figure 24 shows the interior of one version of the multi -analyte breathalyzer device which contains seven sensors. As an example, each sensor can detect each of the following gases: 2- propanol, acrylonitrile, carbon disulfide, dimethyl sulfide, ethanol, isoprene, trimethylamine or any combination of Volatile Organic Compounds (VOCs) which indicate the presence of Inflammatory Bowel Disease (IBD).

[0085] 84. Figure 25 shows a schematic of the interior of a two-sensor device.

[0086] 85. Figure 26 shows a schematic of the interior of a seven-sensor device.

[0087] 86. Figure 27 shows the screen of a multi-sensor, multi-analyte breathalyzer device with a drop-down menu and the various diseases the breathalyzer of this embodiment can test for.

[0088] 87. Figure 28 shows the high sensitivity of PANI / DNNSA to two concentration of hydrogen gas, 20kppm and 6kppm

[0089] 88. Figure 29 shows Principal Component Analysis (PCA) of data from breath testing for hydrogen in breath of individuals with suspected Celiac disease. The PCA method demonstrates clustering of the positive for Celiac disease breath tests. 89. Figure 30 shows the high sensitivity of SPANI to CO2

[0090] 90. Figure 31 shows capability of a breathalyzer device to analyze data from breath testing using artificial intelligence-machine learning. Machine learning algorithms filed within the Arduino microprocessor of a breathalyzer device derive the results with better accuracy than PCA or standard statistical analysis.

[0091] 91. Figure 32 shows capabilities of machine learning algorithms to better analyze the data obtained by using a breathalyzer device for breath testing.

[0092] 92. Figure 33 shows 100% accuracy of identifying the correct diagnosis among users of breathalyzer device who are considered to have symptoms compatible with a certain disease.

[0093] 93. Figure 34 shows the location for the intervention (steps 6 and 7) and insertion of learned file with algorithms in microprocessor to perform analysis of data from user of a breathalyzer device of this embodiment. In step 6, the recalled data pass through a mathematical model which is coded in the microcontroller (fitting) and yield the multiple key parameters. The key parameters along with the user's input information (age, body weight, height, GI related survey, and etc) pass through a trained Machine Learning model or other classification model which was generated by KNN (20) patients’ data and implanted in the microcontroller and return the classification result.

[0094] 94. Figure 35 shows a schematic of a circular tubular channel with multiple sensors on its periphery. The breath is inserted through a sealing opening of the channel where the mouthpiece is securely attached.

[0095] 95. Figures 5 and 34: In step 6 of the device operational flow (Figs. 5 and 34), the recalled data are passed through a mathematical model code programmed into the microcontroller for calibration and (using this information) for calculation of the result. In step 7, after calibration and calculation of data, the result is displayed on the LED screen. In step 6 , the (Figs. 5 and 34) recalled data pass through a mathematical model which is coded in the microcontroller (fitting) and yield the multiple key parameters. The key parameters along with the user's input information (body weight, height, GI related survey, and etc) pass through a trained Machine Learning model or other classification model which was generated by KNN (20) patients’ data and implanted in the microcontroller and return the classification result. 96. Figure 36 shows 3-D drawings of a multisensor breathalyzer device with the removable and replaceable mouthpiece insertion on the side of the device. The mouthpiece is inserted in the opening located on the main body where the sensors are located. Graphic A demonstrates the channel through which the breath sample comes in contact with the sensors. Graphic B demonstrates the location of the sensors in the main body. Graphic C demonstrates the side of the device where the opening for the insertion of the mouthpiece is located. Desiccant, if required, will be placed within the sensor compartment of the main body of the device.

[0096] 97. Figure 37 shows 3-D drawings of a multisensor breathalyzer device with the removable and replaceable mouthpiece insertion in the center of the portion of the main body where the sensors are located. Graphic A demonstrates the location of the sensors and the opening for the exit of the breath. Graphic B demonstrates the main body of the breathalyzer device with the mouthpiece in the center of the compartment where the sensors are located within the main body. Graphic C demonstrates the opening for the insertion of the mouthpiece in the front of the main body within the compartment where the sensors are located.

[0097] 98. Figure 38 shows a schematic of one-sensor or multi-sensor wearable device. The device is a 2-ply face mask (parts A and B schematics). The 2-ply mask, which can have a third ply, if needed, is made of material which allows breath to pass through by diffusion or other direct method and does not permit the return to the mouth and nose. Part A houses one or more than one sensor which includes material selective to gas or gases in breath. Part B houses Al-enhanced software and hardware to operate the device. The sensors are connected, through the connectors, to the processor. The device can include desiccant, as needed. The desiccant crystals or other type of filter are located in the Part A of the wearable device when the sensors are housed.

[0098] 99. Figure 39 shows a schematic of a miniaturized wearable one-sensor or multi-sensor breathalyzer device, worn over the wrist (C) or another part of the body. Schematic A shows the exterior top of the wearable device. The top includes the opening for the insertion of the removable and replaceable mouthpiece, the screen and the on / off switch. The opening for the mouthpiece has a cover which opens to have the mouthpiece securely placed in the opening and after the breath testing process is completed the cover securely closes the opening. Schematic B shows the bottom and inside of the wearable breathalyzer device. The inside includes one or more gas-selective sensors, desiccant, connectors for the sensors to connect to the processor, the processor and the electrical circuit. When the removable and replaceable mouthpiece is in place, the tip of the mouthpiece protrudes into the bottom piece of the wearable breathalyzer. Schematic D shows another embodiment of the configuration of the inside of the wearable breathalyzer. In this configuration the mouthpiece opening is in the center of sensor compartment configuration. The opening for the exit of breath after it comes in contact with the sensors is on the side as depicted by the small arrow in schematic D.

[0099] DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS

[0100] 100. The following detailed description is to be read with reference to the drawings, in which like elements in different drawings have like reference numerals. The drawings, which are not necessarily to scale, depict selected embodiments and are not intended to limit the scope of the invention. Skilled artisans will recognize that the examples provided herein have many useful alternatives that fall within the scope of the invention.

[0101] 101. In certain embodiments, the present invention provides an improved breath analyzer and breath test method to determine the presence of H. pylori in a subject’s digestive tract. The improved breath analyzer and breath test is less costly, more convenient, and more diagnostically accurate than existing methods and devices.

[0102] 102. Exhaled human breath may contain 100 times more CO2 than inhaled air. Exhaled CO2 comes from various sources within the human body. One of these sources can be the presence of H. Pylori in the gastrointestinal tract. Exhaled human breath contains about 3.8% CO2 in healthy individuals who are not infected with H. Pylori.13C is the naturally occurring isotope of elemental carbon.12C is the more stable isotope of carbon-12 and is in CO2.12C exists in nature in abundance at 98.9% of the amount of element carbon.13C, which is present in13CO2, is less abundant in nature and consists of only 1.1% of the natural element carbon.

[0103] 103. H. Pylori is a genotypically diverse bacterium that has the capacity to change its genetic makeup and mutate in vivo during colonization within a human subject’s digestive tract. When H. Pylori is positive for the cytotoxin-associated gene A (CagA), the risk for development of stomach cancer increases relative to when PI. Pylori is negative for CagA. In the western world, H. Pylori seropositivity for CagA is approximately 60% as opposed to the Asian countries and most of Africa where the seropositivity for CagA approaches 100% within the H. Ey / orz-affected population. The highly immunogenic CagA protein encoded by the CagA gene elicits serum antibody responses which can be detected by enzyme-linked immunosorbent assay (ELISA).

[0104] 104. Despite differences in genetic makeup, all types of H pylori are able to hydrolyze urea (either13C labeled or unlabeled) using its abundant enzyme urease to produce CO2 (or13CC>2) and ammonia (NH3) gas. Once produced, ammonia and carbon dioxide (CO2 or13CO2) are diffused in the bloodstream through the gastrointestinal mucosa and exhaled from the lungs through exhaled breath. The hydrolysis of urea, both in labeled and unlabeled form, is shown by Equations 1 and 2 below:

[0105] 105. Equation 1 :13CO(NH2)2+ HOH — urease >13CO2+ 2NH3

[0106] (13C labeled urea) (ammonia)

[0107] 106. Equation 2: CO(NH2)2+ HOH — urease— > CO2+ 2NHi

[0108] (unlabeled urea) (ammonia)

[0109] 107. Prior urea breath tests require labeled urea substrates to detect the amount of labeled CO2 in the breath. In contrast, the present breath analyzer and breath test method uses one or more sensors to detect the total amount of both ammonia and carbon dioxide present in a breath sample. Advantageously, the present breath analyzer and urea breath test method do not require a labeled substrate.

[0110] 108. Referring to the drawings, and in particular, FIGS. 1-11, 21-27 and 34-37, there are shown schematics of types of breath analyzer of the present disclosure. In some embodiments, the breath analyzer includes some or all of the components depicted in these figures. In other embodiments, the breath analyzer includes additional components, other than those depicted in these figures.

[0111] 109. Certain embodiments of the breath analyzer detect presence and concentration of both ammonia and carbon dioxide in a breath sample. As described in greater detail below, the breath analyzer includes an input, one or more gas sensors, an electrical circuit, and a processor. The input receives a breath sample. The at least one gas sensor contacts the breath sample. At least one of the one or more gas sensors includes ammonia selective material (e.g., doped polyaniline) and a conductive material. The electrical circuit operably connects the conductive material to the processor. The processor detects changes in resistivity in the electrical circuit and uses the changes in resistivity to calculate a concentration of ammonia in the breath sample. The same or a different gas sensor includes carbon dioxide selective material (e.g., doped polypyrrole or sulfonated polyaniline (SPANI) and a conductive material. The electrical circuit operably connects this conductive material to the processor. The processor detects changes in resistivity in the electrical circuit and uses the changes in resistivity to calculate a total concentration of carbon dioxide in the breath sample.

[0112] 110. The breath analyzer includes a mouthpiece that has open ends to allow a breath sample to move therethrough. The mouthpiece can comprise any suitable type of material, including, but not limited to, plastic or metal. The mouthpiece includes a first portion and a second portion. The first portion is configured as an input that receives a breath sample. The first portion can be sized and shaped to receive a user's lips, so that a user can blow exhaled breath into the mouthpiece.

[0113] 111. In some cases, the mouthpiece includes a one-way valve. In such cases, a user blows exhaled breath into the mouthpiece. The exhaled breath moves forward pass the one-way valve and becomes trapped. In other words, the exhaled breath cannot move backward past the one-way valve and toward the first portion.

[0114] 112. The breath analyzer also includes a main body. The mouthpiece is attached to the main body. In particular, the second portion of the mouthpiece is sized and shaped to removably connect to the main body. For example, the second portion can be snapped onto or perhaps screwed onto the main body. The main body can comprise plastic, metal, or any other suitable material. In some cases, the main body and the mouthpiece comprise the same material. In other cases, the main body and the mouthpiece comprise different materials.

[0115] 113. In certain embodiments, the mouthpiece is a single-use mouthpiece. A single-use mouthpiece is desirable because it can be replaced for use with each new user. Also, in some cases, components of the mouthpiece and / or main body in contact with exhaled breath can be made of an inert or non-reactive material (e.g., polytetrafluoroethylene (PTFE)) that does not interfere with ammonia absorption.

[0116] 114. The mouthpiece can be integral with the main body, or can be a separate structure that is connected to the main body. In instances where the mouthpiece is a separate structure connected to the main body, the mouthpiece can be placed inside the main body through a hole in the main body using, for example, a push-in, screw-in, or tack-in motion.

[0117] 115. In one embodiment, the mouthpiece is permanently attached to the main body. In such instances, the mouthpiece can be securely mounted on the main body, extending straight out from the main body or at an angle from the main body. These alternate configurations allow the eyes of the subject taking the breath sample to either directly face the main body or to face away from main body while taking the breath sample. The mouthpiece can be permanently mounted to an opening in the main body using a receptacle made of plastic or metal or any other material. In other cases, the mouthpiece can be permanently attached to the main body without the use of a receptacle.

[0118] 116. FIGS. 1 and 21 illustrate exemplary embodiments of the breath analyzer. FIG. 1 shows the breath analyzer and the receptacle or position of the mouthpiece to be attached or screwed on to the main body, whereas FIG. 21 shows the breath analyzer with the mouthpiece being part of the main body. The breath analyzer in these embodiments is provided as a self-contained, portable hand-held device. In some cases, the mouthpiece is a single use mouthpiece that is disposed of after use, and that can be replaced with a new mouthpiece for each new user.

[0119] 117. The mouthpiece can be attached to an exterior of the main body or can extend into the main body of the breath analyzer. The mouthpiece can be attached anywhere on or within the breath analyzer, provided that a first end of the mouthpiece is accessible to lips of the subject undergoing the breath test. The mouthpiece can attach to the breath analyzer via any suitable type of connection, including a straight connection, push-in connection, or screw-in connection, or have another type of connection within the main body of the breath analyzer or be an opening of the body of the breath analyzer. In some cases, the mouthpiece can be glued or can use any other type of adhesive to adhere the mouthpiece to the main body.

[0120] 118. The mouthpiece can have any desired shape. For example, the mouthpiece can be oblong, cylindrical, cone-shaped, or straw-shaped. The shape of the mouthpiece should be such that the lips of the subject are able to wrap around the mouthpiece in a tight manner. As disclosed above, the mouthpiece can optionally include a self-sealing, one-way valve to seal the breath sample from the surrounding air once the breath sample exits the mouthpiece and enters the main body of the breath analyzer. 119. The mouthpiece can optionally include a lining material positioned inside of the mouthpiece. Where provided, the lining material covers some or all of an interior surface of the mouthpiece. In some cases, the lining material is a desiccant that can trap humidity. The desiccant can comprise (consist of, or consist essentially of) silica, activated charcoal, calcium sulfate, calcium chloride, or any other type of desiccant. The desiccant can also include a combination of any one or more of these or other desiccants. The desiccant can optionally have a color indicator to indicate the amount of humidity that the desiccant has trapped. In other instances, the mouthpiece is devoid of any type of desiccant or other lining material.

[0121] 120. In another embodiment, the mouthpiece includes a lining material configured to trap and absorb some of the gases contained in a breath sample, while allowing other gases to go through. As an example, the lining material on the mouthpiece can block H2O, nitric oxide (NO), methane gas (CH4), nitrogen gas (N2), and other volatile organic compounds contained in human breath, while allowing CO2 (or13CO2) and NH3 or another gas contained in the human breath to pass through the lining material and through the mouthpiece.

[0122] 121. The breath analyzer can optionally include one or more filters (or “traps, or sieves”). The one or more filters perform a similar function as the lining material for the mouthpiece. That is, the one or more filters will allow certain gases to pass through, while trapping other gases and preventing them from passing through. The material for the one or more filters can comprise sodium hydroxide, silica, activated charcoal, calcium sulfate, calcium chloride, or any other type of desiccant. The filter can also include a humidity sensor, such as a hygrometer. However, in some cases, the breath analyzer does not include any filters.

[0123] 122. In one embodiment, the filter is a single filter configured to block the passage of certain gases that are present in human breath and that are not intended to be measured by the breath analyzer. As an example, the filter can block humidity, nitric oxide, methane, oxygen gas, and / or other volatile organic compounds present in a breath sample. This single filter allows the passage of ammonia (NH3) and CO2 and13CC>2 or only ammonia or only CO2 or only13CC>2.

[0124] 123. In another embodiment, the filter comprises multiple filters. The filters can have any desired shape and can comprise various types of materials. The shape of the filters is not limiting, and can be round, oblong, square or a combination of different shapes. Each of these filters can trap one or more of the undesirable gases (i.e., those gases not intended to be measured), and allow CO2,13CO2and ammonia to pass through, either separately or together.

[0125] 124. The breath analyzer includes at least one gas sensor, a processor and a power source. The at least one sensor, processor and power source are electrically connected via an electrical circuit.

[0126] 125. The processor can be any desired processor known in the art. In some cases, the processor is a microcontroller. In certain cases, the processor is an Arduino microcontroller. In some cases, insertion of learned fde with algorithms in microprocessor to perform analysis of data from user of the breathalyzer device of this embodiment. The recalled data (Fig. 34) pass through a mathematical model which is coded in the microcontroller (fitting) and yield the multiple key parameters. The key parameters along with the user's input information (age, body weight, height, symptoms, etc.) pass through a trained Machine Learning model or other classification model which is generated by algorithms based on patients’ data and implanted in the microcontroller and return the classification result.

[0127] 126. The at least one sensor can have any of the embodiments already described. In some cases, the at least one sensor includes the sensor shown in the embodiment of FIG. 9. The at least one gas sensor is electrically connected to the electrical circuit using any desired connection mechanism. In some cases, the at least one sensor connects to the electrical circuit via an optional sensor mount (FIG. 15). In such cases, the at least one sensor can be mounted directly onto the sensor mount. The sensor mount serves as an interface between the sensor and the electrical circuit. Thus, the sensor mount can be any structure known in the art that connects the electrodes of the at least one sensor to the electrical circuit. In some embodiments, the sensor mount is a printed circuit board.

[0128] 127. In other cases, the at least one sensor is directly connected to the electrical circuit. For example, in some embodiments, the electrical circuit includes two metal clips that can be clamped onto the contact pads to create an electrical connection. A user can also replace an old sensor with a new sensor by pulling the old sensor out of the metal clips and inserting a new sensor into the clips.

[0129] 128. The main body also includes an on / off button or switch and the power source. The power source can be a portable power source, such as a battery. When the on / off button is activated, the power source turns on. The power source supplies voltage to a voltage regulator. In certain cases, the power source supplies volts to the voltage regulator. The voltage regulator regulates the amount of voltage sent to the sensor. In some cases, the voltage regulator supplies a voltage to the at least one sensor in the amount of between 0 volts to 5 volts. In certain cases, the voltage regulator supplies a voltage to the at least one sensor in the amount of about 5 volts. In one embodiment, the voltage regulator is an IC1 7805 voltage regulator, a product manufactured by Fairchild Electronics.

[0130] 129. A resistor is also electrically connected to the at least one sensor and provides resistance to the at least one sensor. In some case, the resistor is a 10 kQ resistor. When the breath sample contacts the at least one sensor, a change in resistivity occurs in the sensor that correlates to an amount of ammonia and / or amount of carbon dioxide present in the sample, and / or an amount of another gas contained in the human breath. The at least one sensor outputs voltage (along with the changes in resistivity) to the processor. The processor detects changes in resistivity in the at least one sensor and uses the changes in resistivity to calculate a concentration of ammonia and carbon dioxide in the breath sample. The processor can also compare a concentration of ammonia between two different breath samples. Similarly, the processor can compare a concentration of carbon dioxide between two different breath samples.

[0131] 130. The gas sensor can comprise a single gas sensor or more than one gas sensor. Each gas sensor is configured to detect and measure one or more of ammonia, CO2, and13CO2 in a human breath sample. Thus, at least one gas sensor is sensitive to ammonia (NH3), at least one gas sensor is sensitive to CO2, and at least one gas sensor is sensitive to13CC>2. For example, one gas sensor can be sensitive to ammonia, a different gas sensor can be sensitive to CO2, and yet a different gas sensor can be sensitive to13CO2. Alternatively, a single gas sensor can be sensitive to any combination of these gases. As an example, one gas sensor can be sensitive to ammonia and to CO2 and to13CO2; another gas sensor can be sensitive to ammonia (and not to CO2 or13CO2); and another gas sensor can be sensitive to CO2 and to13CO2 (and not to ammonia). One or more of gas sensors can measure the humidity in breath to assess its effect on the sensitivity of the gas sensor to the particular gas under detection.

[0132] 131. In certain embodiment the sensor can comprise more than one sensor or six sensors, or seven sensors or more than twenty-one (21) sensors sensitive to the same number of gases contained in the human breath. Or in another embodiment one sensor can be sensitive, simultaneously, to more than one gases contained in the human breath. As an example one sensor can be sensitive to hydrogen and to ammonia.

[0133] 132. In certain embodiments the sensor can comprise of isoprene-elective material. The material can be doped polyaniline. In other embodiments the material can be thin films of metal oxide composite with nanoparticles, e.g., ZnO thin films. In other embodiments the material can be polymer composite with graphene. In certain embodiments, the concentration of isoprene is in the range of 5-500ppb. In other embodiments isoprene is detected below 5ppb and above 500ppb.

[0134] 133. In certain embodiments the sensor can comprise of oxygen -elective material . The material can be doped polymer, e.g., polyaniline doped with CSA. In other embodiments the material can be polymer composite with graphene or metal oxide composite with nanoparticles.

[0135] 134. In certain embodiments the sensor can comprise of nitrogen-selective material. The material can be doped polyaniline, or metal oxide composite with nanoparticles.

[0136] 135. The breath analyzer can measure ammonia and CO2 (including12CC>2,13CC>2, and / or both12CO2and13CO2) either simultaneously or in succession. Where the breath analyzer measures ammonia and CChin succession, the measurements occur within a short period of time (e.g., from about 1-10 seconds) of each other. This enables the breath analyzer to measure and process the effect of each gas sensor simultaneously or nearly (i.e., substantially) simultaneously.

[0137] 136. The breath analyzer can measure hydrogen and methane either simultaneously or in succession. Where the breath analyzer measures hydrogen and methane in succession, the measurements occur within a short period of time (e g., from about 1-10 seconds) of each other. This enables the breath analyzer to measure and process the effect of each gas sensor simultaneously or nearly (i.e., substantially) simultaneously.

[0138] 137. The breath analyzer can measure hydrogen, methane, ammonia, CO2, acetone, and ethanol simultaneously or in succession. Where the breath analyzer measures hydrogen, methane, ammonia, CO2, acetone, and ethanol in succession, the measurements occur within a short period of time (e.g., from about 1-10 seconds) of each other. This enables the breath analyzer to measure and process the effect of each gas sensor simultaneously or nearly (i.e., substantially) simultaneously. 138. The breath analyzer can measure nitrogen, oxygen, CO2, methane, acetone, isoprene and methanol simultaneously or in succession. Where the breath analyzer measures nitrogen, oxygen, CO2, acetone, isoprene and methanol in succession, the measurements occur within a short period of time (e.g., from about 1-10 seconds) of each other. This enables the breath analyzer to measure and process the effect of each gas sensor simultaneously or nearly (i.e., substantially) simultaneously.

[0139] 139. The breath analyzer can be an autonomous wearable device which is worn over the face. The over the face wearable can be a single-ply or two-ply or multi-ply face mask which includes one or more than gas-selective sensor, software and hardware and power source and a removable mouthpiece. The wearable device can be worn over the mouth and nose or over the mouth or over the nose. In some embodiments the wearable device can be worn over the wrist, (e.g., as a watch or part of a watch) and include one or more than one sensor, a removable mouthpiece, software and hardware. In other embodiments the wearable device can be attachable band (e.g., Band-Aid) placed near or at the base of the nose and contain one or more than one sensor, software and hardware. The software and hardware will collect the data, analyze the data, store the data and transmit the results to a smart phone, computer or the Cloud.

[0140] 140. Each gas sensor includes a substrate, an electrically-conductive material, and a gas- selective material. The electrically-conductive material is deposited onto the substrate. The electrically-conductive material can comprise (consist of, or consist essentially of) any desired electrically-conductive material. In some cases, the electrically-conductive material is platinum. In other cases, the electrically-conductive material is gold. In other cases, the electric conductive material is absent and the gas selective material is placed directly on the surface within the body of the device. As an example, the gas selective material can be placed on material of a 2-ply facial mask which serves as wearable multi-sensor device.

[0141] 141. In certain embodiments the sensor includes a gas-selective material but does not require an electrically-conductive material. As an example, a sensor made of doped PANI / DNNSA does not require to be placed on finger electrodes and PANI / DNNSA is placed directly on the mount which is then connected to the electrical circuit.

[0142] 142. In certain embodiments, the electrically-conductive material is an electrode arrangement. The electrode arrangement can be a single electrode or a plurality of electrodes. The electrodes can be spaced apart in any desired arrangement. In some cases, the electrodes are spaced less than about 250 pm apart, perhaps less than about 150 pm apart, such as about 100 pm apart. In certain cases, the electrodes are spaced less than about 10 pm apart, such as 5 pm apart. In some embodiments, such as the gas sensor shown in FIG. 13, the electrodes include interdigitated finger electrodes. In one embodiment, the gas sensor comprises interdigitated platinum finger electrodes with a line spacing of 100 pm apart or less.

[0143] 143. The one or more gas sensors can operate at room temperature (i.e,. 68-77°F, or 20-25°C) or at a higher temperature. The one or more gas sensors can be fabricated by any suitable method. Such methods can include spin coating, drop-coating, sol-gel, or any other known fabrication method. The one or more gas sensors can be fabricated for either single use or for multiple, repeated uses. The one or more gas sensors can comprise polyaniline or polypyrrole doped with a protonic acid. As non-limiting examples, the one or more gas sensors can comprise PANI / CSA (polyaniline / camphorsulfonic acid); PANI / DNNSA (polyaniline / dinonylnapthalenesulfonic acid); or PPY / DBSA (polypyrrol e / dodecylbenzynesulfonic acid); or SPANI (sulfonated polyaniline). Applicant has found that a PANI / CSA gas sensor can be sensitive to detecting ammonia, and further that PANI / DNNSA and PPY / DBSA can be particularly sensitive to detecting H2,12CC>2 and / or13CC>2.

[0144] 144. The one or more gas sensors can comprise thin-fdm or thick-film sensors capable of sensing ammonia gas (NH3) and / or carbon dioxide gas (CO2 and / or13CO2) simultaneously or in succession. Such gas sensor or sensors can comprise polymer-thin film or polymer-thick film or polymer composite film. The one or more gas sensors can be metal oxide sensors such as zinc oxide (ZnO) or other metal oxide sensors. The one or more gas sensors can be a blend of polymer with metal oxide (such as polyaniline, polypyrrole, or another polymer, in combination with zinc oxide, or another metal oxide). In some cases, the one or more gas sensors comprise sulfonated polyaniline or polyethylenimine (PEI) blended with polyelectrolytes. In other cases, the one or more gas sensors can be emeraldine-base polyaniline (EB-PANI) blended with poly(3,4- ethylenedioxythiophene) polystyrene sulfonate (PEDOT:PSS). In other cases, the one or more gas sensors can be other types of polymer-bases sensors.

[0145] 145. In some cases, the ammonia selective material includes doped polyaniline. Polyaniline exhibits three different oxidation states: leucoemeraldine (LEB, fully reduced), emeraldine (EB, half-oxidized), and pernigraniline (PNB, fully oxidized). Also, when polyaniline is in the emeraldine state, it can be in either an emeraldine salt or emeraldine base form. When in the emeraldine salt form, the polyaniline is conducting. The emeraldine salt form is usually obtained by protonating the basic amine and imine sites with strong acids. This process is reversible in that the emeraldine base form is obtained by deprotonating the amine groups. Thus, the emeraldine state of polyaniline transitions between an acid form and base form.

[0146] 146. The inventor has discovered that the acid-base transition of polyaniline renders it pH sensitive and this characteristic allows it to be effectively used in ammonia detection. When ammonia contacts an emeraldine salt form of polyaniline, the ammonia deprotonates the amine groups and converts it to an emeraldine base, which also causes an increase in resistivity and a corresponding decrease in conductivity.

[0147] 147. In some case, the one or more gas sensors can include a dopant to increase sensitivity of the gas sensor. For example, where polyaniline is used for the at least one gas sensor, the polyaniline can be doped with a protonic acid to increase its pH sensitivity. Polyaniline with increased pH sensitivity is desirable for ammonia detection because when ammonia converts the polyaniline to an emeraldine base, it causes an even larger increase in resistivity and corresponding decrease in conductivity. Larger increases in resistivity (and decreases in conductivity) are desirable because they are easier to detect, and thus increase the sensitivity of the polyaniline to ammonia.

[0148] 148. Polyaniline doped with a protonic acid has increased pH sensitivity compared to undoped polyaniline. In some cases, the polyaniline can be doped with a protonic acid including ions such Cl and SO to obtain pH sensitivity of around 59 mV. Certain protonic acids cause an even larger increase in pH sensitivity. For example, polyaniline doped with camphor sulfonic acid has been shown to have a pH sensitivity of around 70 mV.

[0149] 149. In some cases, the polyaniline comprises at least one dopant that increases pH sensitivity of the polyaniline. In some cases, the dopant is a protonic acid. In some embodiments, the dopant is hydrocholoric acid. In other embodiments, the dopant is camphor sulfonic acid. In yet other embodiments, the dopant is both hydrocholoric acid and camphor sulfonic acid. Other possible dopants include, but are not limited to, sulfuric acid, salicylic acid, acetic acid, citric acid, tartaric acid, oxalic acid, malonic acid, succinic acid, glutamic acid, adipic acid and phthalic acid. Also, in some cases, the polyaniline has a dopant that provides the polyaniline with a pH sensitivity of more than 59 mV. In one embodiment, the polyaniline has a camphor sulfonic acid dopant, which provides the polyaniline with a pH sensitivity of about 70 mV, which is a higher sensitivity observed than when using other dopants.

[0150] 150. Also, the gas-selective material of the gas sensor can be deposited directly onto the electrically-conductive material using any desired deposition process. For example, the gas- selective material can be deposited onto the electrically-conductive material using a spin coating method, a drop coating method, a chemical vapor deposition method, or a sputtering method. In certain cases, the electrically-conductive material is coated with spun cast gas-selective material. In certain embodiments, the gas-selective material is doped polyaniline deposited directly onto the electrically-conductive material using a spin coating method.

[0151] 151. As discussed above, the one or more gas sensors can detect and measure one, two, or more than two gases. At least one of the one or more gas sensors detects ammonia gas (NH3) at very low ppb levels. In some cases, the one or more gas sensors 50 detect ammonia concentration levels in a range from 1 ppb up to 1 ppm. In some cases, the one or more gas sensors 50 detect ammonia gas in a concentration range of lOppb and higher (e.g., in a range of from about 10 ppb up to about 50 ppb), or in a concentration range of 500 ppb or higher. In still other cases, the one or more gas sensors 50 detect breath ammonia at levels lower than 50 ppb and as high as 500 ppm.

[0152] 152. As noted above, at least one of the one or more gas sensors detects CO2. The CO2 detected can be12CC>2,13CO2, or the ratio of13CO2:12CO2 In some cases, at least one of the gas sensors 50 detects CO2 (or13CO2) at concentrations in a range of 10ppm-1500ppm and higher (Figure 6). In some cases, the at least one gas sensor detects CO2 at levels as low as ImM / min or 50-100ppm. In addition, in some cases, the at least one gas sensor detects13CC>2 at levels as low as low as ImM / min or 50-100ppm.

[0153] 153. The one or more gas sensors can comprise thin fdm polyaniline (PANI) doped with hydrochloric acid (HC1). Such a sensor can be prepared by chemical oxidative polymerization of aniline in aqueous acidic medium (IM HC1) with ammonium persulfate (APS) as an oxidant. The PANI sensor can be doped with acid dopants other than HC1. In some cases, the one or more gas sensors can comprise thin fdm polyaniline doped with camphorsulfonic acid (PANI-CSA) and further doped with HC1 or another acid dopant. 154. In some cases, the one or more gas sensors can comprise thick film polymer pyrrole. Such sensors can be polymerized in the presence of an oxidant such as FeCh or any other acid. Polymerized pyrrole (polypyrrole) can detect and measure CO2 and13CC>2 at concentration levels of 10 ppm and above.

[0154] 155. In certain instances, the one or more than one gas sensors can comprise a fiber optic sensor capable of sensing ammonia, CO2 and / or13CO2. It is envisioned that other gas sensors 50, other than those specifically mentioned herein, can be used as the one or more gas sensors 50 to detect and measure ammonia, CO2, and13CC>2.

[0155] 156. One non-limiting method for fabricating a PANI-CSA thin-film sensor is the following: The films can be prepared by spin coating the PANI-CSA solution on interdigitated array (IDA) electrodes. Prior to spin coating, the electrodes can be cleaned by rinsing in methanol followed by rinsing with deionized water and drying in a stream of dry nitrogen. PANI films can then be spun cast onto the IDA electrodes by adding 100 pl solutions at 500 RPM. The electrode pads can be cleaned, e.g., with a Q-tip dipped in methanol, to facilitate direct electrical contact with the breath analyzer.

[0156] 157. The breath analyzer further includes contact pads and an electronic circuit. The one or more gas sensors are connected to the electronic circuit via the contact pads. The electronic circuit is electrically connected to the one or more gas sensors. The electronic circuit can function as shown in Figure 5, or can function as shown in Figure 34. The electronic circuit can register multiple successive values for each measured gas detected by the one or more gas sensors. In some cases, the electronic circuit will register the highest detected value or the average value for each gas detected by the one or more gas sensors. Through the use of software, the electronic circuit will convert the measured resistivity of the gases into concentrations (ppb for ammonia and ppm for13CO2and CO2).

[0157] 158. In some embodiments, the breathalyzer includes a display. The display is electrically connected to the processor. The display is configured to visually display the amount of ammonia, CO2, and13CO2gas detected by the one or more gas sensors. The display can be a window display provided on the main body of the breath analyzer. In an alternative embodiment, the results can be displayed, with the use of Bluetooth technology or any other wireless data transmitter, through a computer portal or other device that can be either stationary or portable. 159. The display can display a single result that represents the additive result of all three gases (ammonia,13CO2 and CO2) or can display one result for each respective gas (i.e., one output for ammonia, one output for CO2, and one output for13CC>2). In some embodiments, the results are displayed in numerical values (e.g., from 1-1000, where a value of 1 represents no H. Pylori infection, and a value of 1000 represents a subject infected with H. Pylori') and / or in actual units of measured gases (e.g., ammonia concentration in ppb; and CO2 and / or13CO2 concentration in ppm). In other embodiments, the results can be displayed as a yes or no indication, and / or in color using a light source. For example, green light on the display can represent a negative test result and red light on the display can represent a positive test result. Any one or more of these results (e.g., numerical values, actual units of measured gases, color, yes / no indication) can be displayed on the display.

[0158] 160. In some embodiments, the main body is configured as including a first compartment and a second compartment. In some cases, as shown in FIGs. 6 and 7, the first compartment is a chamber that houses the at least one gas sensor and a sensor mount, and the second compartment is an electrical housing that houses various components. In certain embodiments (not shown), a separate chamber can be provided for each of the at least one gas sensor.

[0159] 161. In some cases, the chamber includes a lid or door that opens and shuts. When the door is closed, the chamber provides a closed, sealed environment around the at least one sensor. When the door is open, the at least one gas sensor is accessible through the door opening. A user can open the chamber door to remove and replace the at least one sensor as needed. The chamber also includes an outlet. The outlet includes a cap that can be opened to release a breath sample from the chamber and closed to trap a breath sample within the chamber.

[0160] 162. The mouthpiece is connected to the chamber such that exhaled breath passes from the mouth to the mouthpiece directly into the sensor through a channel (figures 6,7, 36, 37) which accepts the exhaled breath from the mouthpiece. In certain embodiments there is one channel; in other embodiments there are two channels. In some embodiments the one channel or more than one channel is configured tube-like; in other embodiments the channeled is configured circularlike (figure 35) or in other types of configuration (Fig. 36 and 37) as long as the channel or channels are configured to reach the sensor or sensors. 163. As discussed above, the breath analyzer can include a fdter. In some cases, the filter is a desiccant assembly. The desiccant assembly helps to remove excess moisture from the exhaled breath. In some embodiments, the desiccant assembly is provided inside of the mouthpiece. In other embodiments, the desiccant assembly is provided inside of the chamber. Exhaled breath first moves through the desiccant assembly before coming into contact with the at least one gas sensor.

[0161] 164. In some cases, the desiccant assembly is provided as a tube upon which exhaled air flows through. The tube can have an interior filled with a plurality of desiccant beads. The desiccant beads can also be arranged such that a plurality of channels are created for exhaled air to flow through. FIG. 22 shows another exemplary embodiment of a desiccant assembly within the breathalyzer. In other embodiments, the filter can have an interior filled with a plurality of desiccant beads arranged such that a single channel is created.

[0162] 165. As discussed above, the breath analyzer can include one or more optional gas filters. The gas filter(s) help to remove a selected gas (e.g., nitrogen and / or hydrogen) from the exhaled breath. In some embodiments, the gas filter is provided inside of the mouthpiece. In other embodiments, the gas filter is provided inside of the chamber. Exhaled breath first moves through the gas filter before coming into contact with the sensor.

[0163] 166. Referring back to FIG. 1,4, 5, 6, 7, 21, 22, 36, 37 the main body can also include a second compartment configured as an electrical housing that houses various components. In the illustrated embodiment, the electrical housing houses the processor, parts of the electrical circuit, a power source, a display, and a wireless connector.

[0164] 167. During use, a user turns the breath analyzer on by activating the on / off switch. The on / off switch can be located anywhere about an exterior surface of the main body. This on / off switch in turn prompts the power source to supply voltage to the voltage regulator. The voltage regulator regulates and supplies voltage to the at least one gas sensor. The resistor also supplies resistance to the at least one gas sensor.

[0165] 168. A user then blows a breath sample into the first portion of the mouthpiece. The breath sample moves past the one-way valve and through the optional desiccant / gas filter. The breath sample then moves out of the optional desiccant / gas filter and into the chamber where it contacts the at least one gas sensor. The breath sample causes a change in resistivity to occur in the at least one gas sensor. This change in resistivity is outputted to the processor. 169. The present disclosure also provides a urea breath test (“UBT”) method that is an improvement of existing urea breath test methods. This novel UBT uses the breath analyzer described above, which is simple to operate, non-invasive, inexpensive and essentially risk-free. The breath analyzer and related breath test method are available at the point-of-care for self-testing, and the results of the test become available instantaneously.

[0166] 170. To undergo this novel UBT, all subjects who wish to be tested for the presence of H. Pylori infection will need to abstain from taking antibiotics, bismuth, sucralfate, and proton pump inhibitors for two weeks prior to using the breath analyzer and breath test method.

[0167] 171. On the day of the breath test method, the subject will follow the following protocol:

[0168] 172. In one embodiment the subject will abstain from food and drink intake for one hour prior to breathing into the breath analyzer. At the end of the one hour abstinence, the subject will exhale into the mouthpiece for the first time by wrapping his or her lips around the mouthpiece and exhaling breath air into the breath analyzer through the mouthpiece with a prolonged exhalation of approximately 5 seconds of time or over a range of 3-8 seconds of time depending on the age, height and weight of the subject. This provides the baseline exhalation.

[0169] 173. After the first exhalation, the subject will ingest urea in quantity calculated as about 5mg / kg of the subject’s body weight or in standard dose of 300mg or in another standard dose of 125mg or 250mg or in another single dose or multiple doses depending on age and weight and general clinical status of the subject. The urea to be ingested can be in two forms. It can be either labeled urea (13CO (NH2)2) or unlabeled urea (CO (NH2)2). Any dose of urea can be taken in a form of a capsule, which can be swallowed with a small amount of water, or can be contained in a small meal, which is either solid (e.g. pudding) or liquid (e.g. citric juice) and that is ingested orally.

[0170] 174. In a span of ten to thirty minutes after ingestion of the urea (capsule or urea-containing meal), the subject will exhale for a second time into the device through the mouthpiece. The posturea exhalation through the mouthpiece can be anywhere in a range of from 10 minutes after ingestion up to 30 minutes after ingestion of the urea. For example, the post-urea exhalation can be at 10 minutes after ingestion, at 15 minutes after ingestion, at 20 minutes after ingestion, or at 30 minutes after ingestion of the urea (labeled or unlabeled). In this embodiment, the second exhalation cannot be earlier than 10 minutes after ingestion of the urea (whether in capsule form or as a urea-containing meal), and cannot be later than 100 minutes after ingestion of the urea (whether in capsule form or as a urea-containing meal). Most commonly, the second exhalation will take place at 15-30 minutes after ingestion of the urea or urea containing meal.

[0171] 175. In an alternative embodiment, after the baseline exhalation, the subject will ingest a high- protein meal (e.g. hamburger, or high protein bar or equivalent). In a span of 20 to 120 minutes after ingestion of the high-protein meal, the subject will exhale into the mouthpiece 12 for the second time. The second exhalation can be at 20 minutes after ingestion, 30 minutes after ingestion, 50 minutes after ingestion, 80 minutes after ingestion, and 120 minutes after ingestion of the high- protein meal. Most commonly, the second exhalation will take place at 60 minutes after ingestion of the high-protein meal.

[0172] 176. The exhaled breaths (baseline and post-urea) enter the main body of the breath analyzer through filter (if present), or directly through the mouthpiece where there is no filter. Each breath exits filter (or mouthpiece) and enters the one or more gas sensors. As the breath samples (baseline exhalation and post-urea exhalation) enter the one or more gas sensors of the breath analyzer, the resistivity of the one or more gas sensors changes in relation to the amount of ammonia and / or CO2 and / or13CC>2 in the breath under examination.

[0173] 177. Where the at least one gas sensor is a single gas sensor, the sensor can detect three gases; ammonia, CO2, and13CO2. Where the at least one gas sensor comprises two gas sensors, one of the gas sensors can detect ammonia, and the other of the gas sensors can detect CO2 and / or13CO2. Where the at least one gas sensor comprises three sensors, one gas sensor can detect ammonia, one gas sensor can detect CO2, and another gas sensor can detect13CC>2. Where the at least one gas sensor comprises four gas sensors, one gas sensor can detect ammonia, a different gas sensor can detect CO2, and yet another gas sensor can detect13CC>2. Where the at least one gas sensor comprises four gas sensors, one gas sensor can detect ammonia, another gas sensor can detect CO2, yet another gas sensor can detect13CO2, and still yet another sensor can detect humidity.

[0174] 178. The change in resistivity due to the presence of ammonia in the breath sample is converted to electrical current and then to parts per billion (ppb) of ammonia. Similarly, the change in resistivity due to the presence of CO2 or13CC>2 is converted to electrical current and then to parts per million (ppm). The conversion of delta (post-urea minus baseline) resistivity (AR) to ppb and to ppm and any measurement of combinations of the gases will be done with the use of software developed exclusively for the function of the breath analyzer and the electronic circuit.

[0175] 179. In one embodiment, display will display a numerical value of the concentration of ammonia in ppb and the numerical value of CO2 (and / or13CO2 and / or13CC>2: CO2) in ppm. In this embodiment, these numeral values corresponding to the gas concentrations will display through a window on the main body of the breath analyzer.

[0176] 180. In another embodiment, the numerical value on display 30 will be a single value derived from calculation pertaining to the amount of ammonia plus the amount of CO2 or13CC>2 or13CO2: CO2 in the breath of the subject at baseline exhalation and at post-urea exhalation. This calculation can be based on equation for linear predictor of outcome. As an example, the equation can have the form:

[0177] 181. (loglO ammonia x constant for ammonia) + (CO2 x constant for CO2) or

[0178] 182. (loglO ammonia x constant for ammonia) + (13CC>2 x constant for13CO2) or

[0179] 183. (loglO ammonia x constant for ammonia) + (CO2 x constant for CO2) + (13CC>2X constant for13CO2) or

[0180] 184. (loglO ammonia x constant for ammonia) + (13CC>2 : CO2 x constant for13CC>2: CO2) or

[0181] 185. another form of polynomial equation where ammonia is in ppb and13CO2 and CO2 are in ppm and13CO2: CO2 is in numerical value.

[0182] 186. The constant is derived from analysis (e.g. regression) of multiple values of ammonia, CO2, and13CO2within the various populations at risk for infection with H. Pylori and populations in general and within the various dietary habits of such populations and populations in general. The equation will be incorporated into software.

[0183] 187. The equation for linear predictor of outcome can have the basic form:

[0184] 188. f(i) =Po+P ixii+ .. .+PpXip for data point i.

[0185] 189. Below a certain number derived from this equation the subj ect will be found to be negative for the presence of infection with H. Pylori. Above a certain number derived from this equation, the subject will be found to be positive for the presence of active infection with H. Pylori. A small number of inconclusive results will be referred for a repeat of the test and if the results remain inconclusive, the subject will be referred for further evaluation through additional means.

[0186] 190. As discussed above, CO2 (or13CO2) and ammonia, which are produced through hydrolysis of urea (unlabeled or labeled) by the H. Pylori-^ oAwcQ enzyme urease are diffused in the bloodstream through the mucosa and exhaled from the lungs through the exhaled breath. Consumption of urea, either labeled or unlabeled, by an infected subject undergoing testing using this invention will produce elevated amounts of CO2 (or13CC>2) and ammonia in the subject’s breath.

[0187] 191. As also discussed above, the subject undergoing testing using this method will exhale into the breath analyzer twice. The first (baseline) exhalation being after one hour abstinence of food and water, and the second (post-urea) exhalation being after the consumption of urea (labeled or unlabeled) or a high-protein meal or high-protein bar. At all times during the method, the breath analyzer 10 will detect and measure the amount of ammonia, CO2 and / or13CC>2.

[0188] 192. When ammonia, CO2 and / or13CO2 levels at post-urea exhalation are higher than at baseline, at a level greater than 100% above baseline levels, the subject being tested can be diagnosed as being infected with H. Pylori. In addition, when the combined ammonia and CO2 (and / or13CO2) levels at post-urea exhalation are at a level greater than 50% above the baseline levels, the subject can be diagnosed as being infected with H. Pylori.

[0189] 193. In diagnosing infection with H. Pylori, the sensitivity and specificity of the presently disclosed method is higher than with prior methods. This is because the presently disclosed method detects the combined amount of ammonia and CO2 (and / or13CO2), rather than detecting ammonia independently of CO2 (and / or13CC>2), or CO2 (and / or13CO2) independently of ammonia. Because the present method measures all products of urea hydrolysis (including ammonia,12CC>2, and13CO2), the method of the present disclosure provides the most accurate method for detecting and diagnosing H. Pylori infection. Simultaneous measurement of CO2 (or13CCh) and ammonia in breath significantly minimizes statistical and other errors which can occur when measuring either CO2 (or13CO2) without ammonia, or ammonia without CO2 (or13CC>2).

[0190] 194. The combined increase of ammonia and CO2 (and or13CC>2) in subjects infected with H. Pylori subjects is multiplicative (or exponential), not additive. Thus, even small differences between the baseline exhalation and the post-urea exhalation would be diagnostic of H. Pylori infection hence significantly decreasing false negative results.

[0191] 195. The combined increase of ammonia and CO2 (and or13CC>2) in infected subjects will be more specific and sensitive for H. Pylori infection than existing breath test methods because the combination of ammonia and CO2 (and or13CC>2) is indicative of, and diagnostic, only for H. Pylori infection (and no other infection) in individuals without end stage liver disease, encephalopathy, end stage renal disease, and metabolic disease (e.g. Krebs’s cycle defect, urea cycle defect).

[0192] 196. Certain embodiments provide for autonomous, hand-held breath testing device which includes seven or more sensors, software with embedded machine learning algorithms and a processor and electrical circuit. The electrical circuit connects operably each sensor with the processor and the processor detects each sensor’s resistivity and converts resistivity to the concentration of gas for which each sensor is selective. The software provide for increased accuracy and classification of breath testing data. Artificial intelligence and machine learning algorithms are utilized to better interpret the data and the results of breath testing methods. Machine learning algorithms are utilized to examine clustering of data and detect data with precision in way that is not accomplished by utilizing standard statistical methods. Machinelearning algorithmic methods include but are not limited to K-means, DBSCAN or KNN. Certain embodiments include algorithms to improve interpretation of data derived from anthropometric, physical, psychological, clinical and breath testing of a subject.

[0193] 197. Certain embodiments provide for a breath test method. The breath test method includes a step of providing an autonomous, hand-held breath testing device which includes a removable and replaceable mouthpiece and a main body. The main body includes 6 or more sensors, software embedded with machine learning algorithms and a processor, and an electrical circuit. The first sensor includes a nitrogen selective material which has resistivity which increases as a result of the increased concentration of nitrogen. The second sensor includes a CO2 selective material with resistivity which increases in response to increased concentration of CO2. The third sensor includes oxygen selective material that has resistivity which increases in response to increased concentration of oxygen. The fourth sensor includes isoprene selective material that has a resistivity which increases in response to increased concentration of isoprene. The fifth sensor includes acetone selective material that has a resistivity which increases in response to increased concentration of acetone. The sixth sensor includes methanol selective material that has a resistivity which increases in response to increased concentration of methanol. The seventh sensor includes nitric oxide selective material that has a resistivity which increases in response to increased concentration of nitric oxide. In certain embodiments the breath test method provides for predicting the athletic performance of a subject. The method includes collecting a breath sample from the subject before, during and after exercise. The exercise includes but is not limited to swimming, cycling, running, playing tennis, basketball, baseball, football, soccer and other Olympic sports. The breath sample contacts each of the seven sensors and the breath testing device measures concentration of nitrogen, oxygen, CO2, isoprene, acetone, methanol, nitric oxide, in breath, before, during and after exercise.

[0194] 198. Certain embodiments provide for an autonomous, standalone, hand-held breathalyzer device which includes one or more than one sensors. The sensors are gas selective for breath gases and VOCs which can be found in the breath of individuals afflicted by C. Difficile infection. These gases and VOCs are propan-l-ol, 3 -methylbutanal, ethyl propionate, hexanoic acid, 4-methyl phenol, dodecane, and indole. In other embodiments, addressing the different ribotypes of C. Diff., gas-selective sensors for methanol, p-cresol, dimethylamine and a range sulfur compounds (ethylene sulfide, dimethyl sulfide and methyl thioacetate) as VOCs are used to differentiate the bacterium’s ribotypes.

[0195] 199. Other embodiments provide for an autonomous, standalone, hand-held breathalyzer device which includes on or more than one gas-selective sensors for breath gases and VOCs, which, studies have shown, are increased in individuals who have colorectal cancer. These gases and VOCs are mainly 1 -iodononane and benzene, although other VOCs like Nonanal, 4-Methyl- 2-pentanone, Decanal, 2-Methylbutane, 1,2-Pentadiene, 2-Methylpentane, 3 -Methylpentane, Methylcyclopentane, Cyclohexane, Methylcyclohexane, 1,3-Dimethylbenzene, 4-Methyloctane, 1,4-Dimethylbenzene, 4-methylundecane and trimethyldecane have been noted.

[0196] 200. Certain embodiments provide that nanostructured films of metallic vanadium oxide are deposited on sensor substrates (Pt interdigitated electrodes [IDE] pre-patterned on silica) using spin coating technique. First, 0.3 g as prepared powders are dispersed ultrasonically in 15 mb ethyl alcohol or IPA for 30 min with an appropriate amount of dispersant, and 5 g Nafion (M.W. 90 000-120 000, Aladdin Chemistry Co., Ltd.), and then the mixture is uniformly cast onto a float glass substrate by spin-coating with the speed of 600 r / min for 20 s and then 1000 r / min for 20 s. After removing the liquid by drying in an oven at 80 °C for 60 min, the VO2 nanoparticle-based thermochromic fdm is obtained. This is the method to produce methane-selective material for sensing of the breath gas methane.

[0197] 201. The following non-limiting examples are intended to show how the breath test method and breath analyzer of the present disclosure can be used to detect H. Pylori infection in humans, adults and children.

[0198] 202. EXAMPLE 1 - Simultaneous calculation of the rise of ammonia and of CO2 in breath after ingestion of unlabeled urea (CO(NH2)2 as diagnostic of H. Pylori infection.

[0199] 203. The subject abstains from antibiotics, bismuth, proton pump inhibitors, and sucralfate for two weeks. At the end of the two-week period, the subject abstains from food and drink for one hour. At the end of the one hour fast, the subject exhales into the breath analyzer through the mouthpiece. The exhalation can last from about 2-10 seconds. In another embodiment, the exhalation continues until a characteristic sound or light coming from the main body indicates that a sufficient amount of exhaled breath has entered the main body. By sufficient, it is meant that there is enough of the breath sample to come into contact with the one or more gas sensors.

[0200] 204. When the baseline exhalation enters the main body, the breath sample passes through filter (when it is in place), where gases like NO, CEL, N2, volatile organic compounds, O2, and humidity are blocked from entering the at least one gas sensor. Ammonia and CO2 are not blocked by the filter and are allowed to contact the one or more gas sensors.

[0201] 205. When the breath sample passes to the one or more gas sensors, the resistivity of the one or more gas sensors changes according to the amount of ammonia and CO2 present in the exhaled breath. The resistivity of the at least one gas sensor is converted to ppb of ammonia and ppm of CO2 with the use of the electronic circuit and software. The numerical value of ammonia and CO2 obtained during the first exhalation can be stored in memory.

[0202] 206. At the end of the baseline exhalation, the subject ingests a known quantity (e.g., 125 mg, 250mg, or 300mg, depending on the weight and age of the subject) of unlabeled urea either in tablet or in a small meal or drink. Tn a span of 10 to 120 minutes post ingestion of unlabeled urea, the subject exhales into the mouthpiece for the second time for the post-urea exhalation, following the same procedure as with the first exhalation. The post-urea exhalation takes place most commonly at 20 minutes post ingestion of unlabeled urea. The post-urea exhalation will not take place earlier than 10 minutes post ingestion, or later than 120 minutes post-urea ingestion of unlabeled urea. The values of ammonia in ppb and of CO2 in ppm of the second exhalation are also stored in memory of the breath analyzer device 10.

[0203] 207. The breath analyzer 10 will calculate the final values of ammonia and CO2 as follows:

[0204] 208. Final ammonia- post-urea ammonia minus baseline ammonia

[0205] 209. Final CC>2=post-urea CO2 minus baseline CO2.

[0206] 210. Software can be used to convert the resistivity values to final ammonia concentration in ppb and final CO2 concentration in ppm. In subjects who test positive for H. Pylori infection, the range of values for baseline ammonia would be 20ppb-200ppb and the range for post-urea ammonia would be 80ppb to 600ppb. Because of the overlap of the baseline and post-urea ammonia values, the actual percent change between baseline and post-urea ammonia will be the marker for the infection with H. Pylori. When the percent change between baseline and post-urea ammonia is greater than 200%, the subject is considered positive for H. Pylori infection. When the percent change between baseline and post-urea ammonia is lower than 200%, the subject is negative for H. Pylori infection.

[0207] 211. In subjects who are positive for H. Pylori, the baseline CO2 concentration is typically below 200 ppm, and the post-urea CO2 concentration is typically above 200ppm (e.g., ranging from 200-1000 ppm). When the calculated percent change between baseline and post-urea CO2 is greater than 1000%, the subject is considered positive for H. Pylori infection.

[0208] 212. In one embodiment, the positive test result will be the value of percent change of ammonia above 200%, and the value of the percent change of CO2 above 1000%, in a numerical value, in ppb and in ppm respectively, as determined using device specific software.

[0209] 213. In another embodiment, the final result, which the device’s software will convert into values recognizable by computer program, will be calculated using the equation which takes into account the percent or actual rise of ammonia and the percent or actual rise of CO2 between baseline and post-urea breath samples. The equation can be a logarithmic or statistical equation with the values of two variables (ammonia and CO2) corresponding to the subject being tested, the two constants, and one or more coefficients. The constants and the coefficients are calculated on the basis of demographics and characteristics (e.g. age, gender, race, height and weight) of subjects infected with H. Pylori.

[0210] 214. In another embodiment, the positive H. Pylori result will be a color (e.g. red), and the negative H. Pylori result will be a color (e.g. green). In another embodiment, the positive result for H. Pylori infection will be a plus sign (+) and the negative for H. Pylori will be a minus sign (-)•

[0211] 215. The final result can be displayed through a window of the main body of the breath analyzer. The final result (whether indicating positive or negative, yes / no, different colored lights, or numerical values of ammonia and of CO2) will be displayed through the window for the main body. The breath analyzer will include instructions for the subj ect undergoing testing so as to allow the subject to interpret the displayed results and provide the subject with recommendations for potential further examination by physician or equivalent personnel and potential treatment options.

[0212] 216. EXAMPLE 2 - Simultaneous Calculation of the rise of ammonia and of13CC>2 in breath after ingestion of13C labeled urea as markers for the presence of H. Pylori infection.

[0213] 217. In this example, the subject abstains from antibiotics, bismuth, proton pump inhibitors, and sucralfate for two weeks. At the end of the two-week period, the subject abstains from food and drink for one hour. At the end of the one hour abstinence, the subject exhales into the mouthpiece. The exhalation lasts 2-10 seconds or until a characteristic sound or light coming from the main body of the breath analyzer indicates that a sufficient amount of exhaled breath has entered the main body through the mouthpiece. By sufficient, it is meant that there is enough of the breath sample to come into contact with the one or more gas sensors.

[0214] 218. At the end of exhalation and when the sufficient amount of exhaled breath enters from the mouthpiece into the main body, it passes first through filter (when present) where gases such as NO, CH4, N2, volatile organic compounds, O2, and humidity are blocked from entering the one or more gas sensors. The filter does not block ammonia or13CO2, which gases are allowed to pass through and contact the one or more gas sensors. 219. Tn another embodiment, the exhaled breath enters into the main body from the mouthpiece directly to the one or more gas sensors. The resistivity of the one or more gas sensors changes according to the amount of ammonia and13CC>2 present in the exhaled breath that comes into contact with the one or more gas sensors. The change in resistivity of the one or more gas sensors is converted to ppb of ammonia and to ppm ofl 3CC>2 through the use of the electronic circuit and software. The numerical value of ammonia and13CO2 obtained during the first exhalation are stored in the memory of the electronic circuit.

[0215] 220. At the end of the baseline exhalation, the subject ingests a known quantity (e.g., 125mg or 250mg or 300mg depending on the weight and the age of the subject) of13C labeled urea, either in tablet form or in the form of a small meal or drink. In a span of 10 to 120 minutes post-ingestion of urea, the subject exhales into the mouthpiece for the post-urea exhalation, following the same procedure as with the baseline exhalation. The post-urea exhalation takes place most commonly at 20 minutes post -ingestion of labeled urea. The post-urea exhalation will not take place earlier than 10 minutes post ingestion or later than 120 minutes post ingestion of labeled urea. The concentration values of ammonia in ppb and of13CC>2 in ppm are stored in the memory of the device.

[0216] 221. The device will calculate the final values of ammonia and13CO2 as follows:

[0217] 222. Final ammonia= post-urea ammonia minus baseline ammonia

[0218] 223. Final13CO2= post-urea13CC>2

[0219] 224. The designed software will convert the values of final ammonia to ppb and final13CC>2 to ppm. In subjects who test positive for / / . Pylori infection, the range of values for baseline ammonia is 20ppb-200ppb, and the range for post-urea ammonia is 80ppb to 600ppb. Because of the overlap of the baseline and post-urea ammonia values, the percent change between pre-urea and post-urea ammonia will be the marker for the infection with H. Pylori. When the percent change between pre-urea and post-urea ammonia is greater than 200%, the subject is considered positive for H. Pylori infection. When the percent change between baseline and post-urea ammonia is lower than 200%, the subject is negative for H. Pylori infection. 225. Tn subjects who are positive for H. Pylori infection, the baseline13CO2 concentration is typically below 200 ppm, and the post-urea13CO2 concentration is typically above 200ppm (and can range, e.g., from 200 ppm up to about 1000 ppm).

[0220] 226. In one embodiment, the positive test result will be the value of percent change of ammonia above 200% plus the value of the percent change of13CC>2 above 1000% as a numerical value determined using device specific software.

[0221] 227. In another embodiment, the final result, which the device’s software will convert into values recognizable by computer program, will be calculated using an equation which takes into account the percent rise of ammonia and of13CC>2 between baseline and post-urea breath samples. The equation can be a logarithmic or statistical equation with the values of two variables (ammonia and13CO2) corresponding to the subject being tested and two constants and one or more coefficients.

[0222] 228. In another embodiment, the positive H. Pylori result will be a color (e.g. red), and the negative H. Pylori result will be a color (e.g. green). In yet another embodiment, a positive H. Pylori result will be the plus sign (+) and a negative result for H. Pylori will be a minus sign (-).

[0223] 229. The final result can be displayed through a window on the main body of the device. The final result (e.g., indicating positive or negative, yes / no, different colored lights, or numerical values of ammonia and of 13CO2) will be displayed through the window on the main body. The device will include instructions to allow the subject undergoing testing to interpret the displayed results and provide the subject with recommendations for potential further examination by physician or equivalent personnel and potential treatment options.

[0224] 230. EXAMPLE 3 - Calculation of the rise of the ratio13CO2 / 12CC>2, as delta over baseline (DOB), in human breath at baseline and post-urea (13C labeled Urea) and calculation of hostdependent urea hydrolysis rate as markers for H. Pylori infection.

[0225] 231. In this example, the subject abstains from antibiotics, bismuth, proton pump inhibitors, and sucralfate for two weeks. At the end of the two-week period, the subject abstains from food and drink for one hour. At the end of the one-hour abstinence, the subject exhales into the mouthpiece. The baseline exhalation takes place over 2-10 seconds (or longer), or until a characteristic sound or light coming from the main body of the device indicates that a sufficient amount of exhaled breath has entered the main body through the mouthpiece. By sufficient, it is meant that there is enough of the breath sample to come into contact with the one or more gas sensors. In another embodiment, the subject exhales into the mouthpiece for a short period of time, which can be shorter than 5 seconds and it can be a short burst of exhaled air which enters the device through the mouthpiece. In this embodiment, the short burst of exhaled air would be a sufficient amount of breath to enter the device through the mouthpiece. There could also be multiple (e.g., more than one) short bursts of exhaled air that will enter the breath analyzer through the mouthpiece.

[0226] 232. When the sufficient amount of exhaled breath enters the main body of the device, and passes through the filter, undesirable gases (e.g., NH3, NO, CH4, N2, volatile organic compounds, and / or O2) and humidity are trapped and do not pass through the filter. The filter does not block CO2 gas, which gas then comes into contact with the one or more gas sensors.

[0227] 233. In another embodiment, the device does not contain filter, and the sufficient amount of exhaled breath over 2-10 seconds (or bursts of exhaled breath) enters the device 10 through the mouthpiece and reaches the one or more gas sensors directly (i.e., without passing through a filter). In such instances, none of the gases contained in the exhaled breath are blocked when passing from the mouthpiece to the one or more gas sensors.

[0228] 234. In another embodiment the device contains filter or desiccant for one sensor within one sensor holder (figure 8) but does not contain filter or desiccant for the other sensor in the other sensor holder. In this type of embodiment, the breath travels from the mouthpiece to the channel to the sensor holder at the end of the channel that corresponds to the holder without desiccant or filter; also, the breath, simultaneously, travels to the other sensor or sensors which have desiccant for the other sensor in the other sensor holder.

[0229] 235. The resistivity of the at least one sensor changes according to the amount of CO2 present in the exhaled breath which comes into contact with the at least one sensor. The change in resistivity is converted to ppm of CO2 with the use of the electronic circuit and software. The numerical value in ppm of CO2 obtained during the first exhalation is stored in the memory of the device with the use of software. 236. In another embodiment, the resistivity of the sensor is not converted to ppm. It remains as a measurement of current that is generated by change in resistivity and is stored in the memory of the device after each exhalation (baseline and post-urea).

[0230] 237. At the end of the baseline exhalation, the subject ingests a known quantity (e.g., 125mg or 250mg or 300mg) of13C labeled urea either in capsule or tablet form, or in the form of a small meal or drink. In a span of 10 to 120 minutes post ingestion of13C labeled urea, the subject exhales into the mouthpiece for a second time, following the same procedure as with the baseline exhalation. The post-urea exhalation takes place most commonly at 20 minutes post ingestion of13C labeled urea. The post-urea exhalation will not take place earlier than 10 minutes post ingestion or later than 120 minutes post ingestion of13C labeled urea. After the post-urea exhalation, the values of CO2 in ppm and of13CO2in ppm are stored in the memory of the device.

[0231] 238. The device will then calculate the final values of CO2 and13CO2 as follows:

[0232] 239. Final CO2= post-urea CO2 minus baseline CO2

[0233] 240. Final13CO2= post-urea13CO2minus baseline13CO2.

[0234] 241. In one embodiment the positive test result will be the value of percent change between13CO2 / CO2of the post-urea exhaled breath over the13CO2 / CO2of the baseline exhaled breath sample of greater than a certain number which will be called the cut-off point. Above the cut-off point, the result will be positive for the presence of H. Pylori infection. Below the cut-off point, the result will be negative for H. Pylori infection. The cut-off point will be calculated during the clinical examination of patients with H. Pylori infection undergoing the present breath test. The highest level of13CO2 / CO2demonstrated by the subjects (in clinical trials) who test negative for H. Pylori infection will be the cut-off point. Levels above this cut-off point will indicate positive for H. Pylori infection.

[0235] 242. In another embodiment, the final result will be calculated using an equation which takes into account the percent rise of CO2between baseline and post-urea breath samples. The equation can be a logarithmic or statistical equation with the values of one variable (CO2) corresponding to the metabolic rate of the subject being tested, one or more constants, and one or more coefficients. This equation will be used to discern the range of values above which the results will be positive for H. Pylori infection and below which the results will be negative for H. Pylori infection. 243. Tn another embodiment, the positive H. Pylori result will be a color (e.g. red), and the negative H. Pylori result will be a color (e.g. green). In another embodiment, the positive / . Pylori result will be a plus sign (+) and a negative H. Pylori result will be the minus sign (-).

[0236] 244. The final result will be displayed through a window on the main body 14 of the device 10. The final result (e.g., indicating positive or negative, yes / no, different colored lights, or numerical values of the DOB (Delta over Baseline) for13CO2 / CO2) will be displayed through the window on the main body of the device. The device will include instructions to allow the subject undergoing testing to interpret the displayed results and provide the subject with recommendations for potential further examination by physician or equivalent personnel and potential treatment options.

[0237] 245. Calculation of host-dependent urea hydrolysis rate (UHR):

[0238] 246. UHR = CO2 produced x delta over baseline x 0.3463, where delta over baseline is defined as the difference between baseline CO2 (or13CO2) and post-urea CO2 (or13CC>2).

[0239] 247. In certain embodiments, the present disclosure provides a multi-sensor breath analyzer (breathalyzer) device which analyzes the breath of an individual for the presence of gases including, but not limited to, acetaldehyde, 2-propanol, acetonitrile, acrylonitrile, benzene, isoprene, pentane, methylexane, ethane, hydrogen sulfide, triethyl amine, trimethyl amine, carbon disulfide, dimethyl sulfide, 1 -Heptene, 1 -Octene, 1 -Nonene, 1 -Decene, methane, ethanol, ammonia (NH3), nitric oxide (NO), nitrogen (N2), hydrogen (H2), Oxygen (O2), carbon dioxide (CO2 or13CO2), carbon monoxide (CO), 2,2,4,6,6-pentamethylheptane, 3,6-dimethyldecane, dodecane, 2,3,4-trimethylhexane, 2,6,8-trimethyldecane, tridecane, undecane, tetradecane, as well as other gases.

[0240] 248. In one embodiment, as shown, for example, in Fig. 1, a breathalyzer device is provided that comprises a main body that can be made of a durable and lightweight material. The main body can include a display (e.g., a touch screen display). The display can extend over (and cover) some or all of one side of the body (e.g., with unbreakable glass). The display can allow for input by the user and for output by the device after the device has finished analyzing the breath of the user.

[0241] 249. The main body of breathalyzer (figures 21, 22, 36, 37) can contain multiple sensors each of which is connected independently to an electronic circuit and microprocessor. The electronic circuit which is connected to each sensor through mechanical connection converts the sensor’s resistivity and current to voltage. The microprocessor, which can be Arduino or another type of microprocessor constructed for use by the breathalyzer, analyzes the signal from the sensor and converts it to voltage. Through software, the voltage is converted to units of measurement for each gas to which each sensor is sensitive and selective.

[0242] 250. In one embodiment, the sensors of breathalyzer are electrochemical sensors. Electrochemical sensors have a conductive portion and a substrate portion. The conductive portion can include, but is not limited to, gold, platinum, palladium or a mixture of gold and platinum. The substrate portion can include, but is not limited to, doped polymers such as doped polyaniline, doped polypyrrole and others. Polyaniline, when doped with protonic acids such as dinonaphthalenesulfonic acid (DNNSA), Camphorsulfonic acid (CSA), hydrochloric acid (HCL), sulfosalicylic acid (SSA) and 4-dodecylbenzenesulfonic acid (DBSA), is rendered highly sensitive to gases such as ammonia, hydrogen, nitrogen, methane and hydrogen sulfate. Polypyrrole doped with FeCh, or with Aminobenzenesulfonic acid (ABSA) or with salicylic acid or another protonic acid is rendered sensitive to CO2 and to13CC>2. Electrochemical sensors such as polymer-based sensors (e.g., doped polyaniline and polypyrrole) are more effective and desirable for the breathalyzer because they are stable and operate at room temperature (about 70 degrees Fahrenheit), the temperature at which the present breath analyzer would optimally operate.

[0243] 251. In another embodiment the sensors of breathalyzer are metal oxide nanosensors, such as ZnO, PbO-doped SnCh, which are sensitive to hydrogen, methanol, propanol and acetone. Other metal oxides can be used as the sensor material, either independently or in combination with other conductive material placed on metal finger electrodes (e.g., platinum or gold or both) or on floating gate field effect transistors (FGFET) or on carbon nanotubes or on nanowires.

[0244] 252. In another embodiment, the sensors of the breathalyzer are polymers (e.g., polyaniline and / or polypyrrole) on conductive materials which are chemical sensitive field effect transistors or floating gate field effect transistors (FGFET) or any other field effect transistors (FETs).

[0245] 253. In one embodiment the breathalyzer comprises a removable mouthpiece or an opening on one side of the breathalyzer which is constructed in such manner for the mouth of the user to be wrapped around it. In one embodiment in which the opening of the device serves as mouthpiece (Figs. 22-26), desiccant can be embedded into the wall of the device itself. In another embodiment in which there is a removable and replaceable mouthpiece, desiccant can be embedded in the wall of the removable and replaceable mouthpiece. The desiccant absorbs humidity at the desired level for the operation of the sensors. In addition, or alternatively, a filter can be placed in front of some (or all) sensors in order to block gas potentially interfering with a particular sensor which is tasked to detect another gas. Such filter can be in the form of crystal or like silica crystal.

[0246] 254. The display of breathalyzer, which can be a touch screen display, and which can occupy the entire side of the body of the breathalyzer much like the display of a smart phone, can include a drop-down menu which provides the user with several options for testing. Such options can include, but are not limited to, the following medical diagnoses: Celiac Disease, NCGS, SIBO, IBS, diabetes, asthma, COPD, Hyperammonemia, kidney disease, liver disease, lung disease, lactose intolerance, fructose intolerance and others. The user touches one or more than one medical diagnoses and the breathalyzer, through its software, asks for input of demographics and characteristics or symptoms compatible with the disease desired to be investigated. After this process is completed to the satisfaction of the preprogrammed algorithm, the user is prompted to exhale into the breathalyzer. The breathalyzer performs the predetermined task of analyzing the breath sample and provides the result with general recommendations.

[0247] 255. In one embodiment, the invention provides a breathalyzer device which contains and utilizes 21 or more sensors each of which is sensitive to one gas. These gases can include acetaldehyde, 2-propanol, acetonitrile, acrylonitrile, benzene, isoprene, pentane, methylexane, ethane, hydrogen sulfide, triethyl amine, trimethyl amine, carbon disulfide, dimethyl sulfide, 1 - Heptene, 1 -Octene, 1 -Nonene, 1 -Decene, methane, ethanol, ammonia (NH3), nitric oxide (NO), nitrogen (N2), hydrogen (H2), Oxygen (O2), carbon dioxide (CO2 or13CO2), carbon monoxide (CO), 2,2,4,6,6-pentamethylheptane, 3,6-dimethyldecane, dodecane, 2,3,4-trimethylhexane, 2,6,8- trimethyl decane, tridecane, undecane, tetradecane and H2O. Under this embodiment, the breathalyzer 100 can be used for screening and monitoring of celiac disease, non-celiac gluten sensitivity (NCGS), IBD (ulcerative colitis and Crohn’s disease), IBS, SIBO, lactose intolerance, fructose intolerance, asthma, COPD, liver disease (steatohepatitis, end stage), H. Pylori infection, kidney failure and metabolic disease (diabetes, genetic).

[0248] 256. In another embodiment, the invention provides a breathalyzer which contains 21 or more sensors and electronically utilizes, on command, all or fewer of the sensors each of which is sensitive to one gas. These gases include but are not limited to acetaldehyde, 2-propanol, acetonitrile, acrylonitrile, benzene, isoprene, pentane, methylexane, ethane, hydrogen sulfide, triethyl amine, trimethyl amine, carbon disulfide, dimethyl sulfide, 1-Heptene, 1-Octene, 1- Nonene, 1 -Decene, methane, ethanol, ammonia (NH3), nitric oxide (NO), nitrogen (N2), hydrogen (H2), Oxygen (O2), carbon dioxide (CO2 or13CO2), carbon monoxide (CO), 2, 2, 4,6,6- pentamethylheptane, 3,6-dimethyldecane, dodecane, 2,3,4-trimethylhexane, 2,6,8- trimethyldecane, tridecane, undecane, tetradecane.

[0249] 257. In another embodiment, the invention provides a breathalyzer which contains 21 or more sensors and electronically utilizes, on command, 17 or fewer of the sensors which are sensitive to one gas each. These gases can include, but are not limited to, 2-propanol, acetonitrile, acrylonitrile, benzene, isoprene, pentane, methylexane, ethane, hydrogen sulfide, triethyl amine, trimethyl amine, carbon disulfide, dimethyl sulfide, 1-Heptene, 1-Octene, 1 -Nonene, 1 -Decene.

[0250] 258. In another embodiment, the invention provides a breathalyzer which contains 21 or more sensors and electronically utilizes, on command, 7 or fewer of the sensors which detect one gas each. These gases can include 2-propanol, acrylonitrile, carbon disulfide, dimethyl sulfide, ethanol, isoprene, trimethylamine.

[0251] 259. In another embodiment, the invention provides a breathalyzer which contains 21 or more sensors and electronically utilizes, on command two or fewer sensors which detect one gas each. Under this embodiment one gas or two gases are detected. These gases include but are not limited to hydrogen (H2), nitric oxide (NO), ammonia, acetone, CO2 or13CO2. Under this embodiment, the diseases for which to screen and monitor include but are not limited to celiac disease, NCGS, IBS, diabetes, H. Pylori infection, asthma, and kidney failure.

[0252] 260. In another embodiment the invention provides a breathalyzer device which contains one or two or more than two sensors each of which is sensitive to either one gas or more than one gas.

[0253] 261. In yet another embodiment, the invention provides a breathalyzer device which contains seven or more than seven sensors each of which is sensitive to one gas or more than one gas.

[0254] 262. In one embodiment the mouthpiece of the breathalyzer is an opening on one of the side of the breathalyzer device. In another embodiment, the mouthpiece is a separate cylindrical piece which can be removably attached to the device. 263. Tn one embodiment, the breathalyzer device contains desiccant (e.g., in the form of crystals) to remove a predetermined amount of humidity from the breath sample and / or a fdter to block certain gas or gases from coming into contact with the operating sensors. The crystals are placed either in the interior of the device in the vicinity of the sensor(s) or are placed on or within the wall of the mouthpiece or under the mouthpiece or in the interior of the device.

[0255] 264. The following non-limiting examples illustrate certain methods that can be used with breathalyzer.

[0256] 265. EXAMPLE 4

[0257] 266. The user wishes to examine whether he / she has Celiac disease by using the present breathalyzer, and can do so using the following procedure:

[0258] 267. 1. The user presses the power button on the front of the device and the device responds by giving a greeting on the screen and asks the user for information (e.g., name, height, weight, age, gender) and guides him / her to tap the drop-down menu.

[0259] 268.2. The user selects (e.g., taps) the drop-down menu and finds the celiac / SIBO icon.

[0260] 269.3. The user selects (e.g., taps) the celiac / SIBO icon and then the celiac icon.

[0261] 270.4. The device responds with a process which the user follows in order to prepare for the test.

[0262] 271. A) The preparation which the device gives out states:

[0263] 272. B) Abstain from high amount of carbohydrate for 48 hrs. and take nothing by mouth for 8hrs., overnight, before exhaling once into the device while fasting; and brush your teeth but don’t use mouthwash.

[0264] 273. 5. When the user is ready, he / she turns the device on, selects (e.g., taps) the drop-down menu and selects (e.g., taps) celiac disease.

[0265] 274. 6. The device asks if the user is ready.

[0266] 275. 7. The user selects (e.g., taps) yes (or no if not ready).

[0267] 276. 8. The device asks the user to exhale through the mouthpiece for about 5 seconds and wait. 277. 9. The user exhales for 5 seconds.

[0268] 278. 10. The device which is programmed through software to test for celiac disease by utilizing the hydrogen sensor blocks all other sensors and operates as a single sensor device.

[0269] 279. 11. The device returns a quantitative result to the user for the amount of hydrogen in the user’ s breath sample and offers an assessment of whether the value is within normal limits. The device prompts the user to discuss the finding with his / her healthcare provider.

[0270] 280. EXAMPLE S:

[0271] 281. The user wishes to have a full profile of his / her metabolomics, and can do so by performing the following steps. In this case, the breathalyzer device utilizes all sensors simultaneously:

[0272] 282. 1. The user turns the device on by pressing on the power button.

[0273] 283. 2. The device gives the greeting and asks the user to select (e.g., tap) the drop-down menu.

[0274] 284. 3. From the menu, the user selects (e.g., taps) “total profile.”

[0275] 285. 4. The device responds with the following process for the user to follow:

[0276] 286. A) Asks for information and whether the user has used antibiotics or other types of medications.

[0277] 287. B) Depending on the responses, the device will either ask the user to wait for two weeks or would recommend overnight fasting (8hrs.) and return for the test in the morning.

[0278] 288. 5. When the user is ready in the morning, he / she has to brush teeth but not use mouthwash.

[0279] 289. 6. The user turns on the device.

[0280] 290. 7. The user selects (e.g., taps) the menu.

[0281] 291. 8. The user selects (e.g., taps) “total profile”

[0282] 292. 9. The device asks the user to exhale through the mouthpiece for 5 seconds and then wait.

[0283] 293. 10. The user exhales for 5 seconds. 294. 11. The device utilizes all sensors simultaneously and responds with readout for all detected gases and offers an assessment of whether the values are within normal limits. The device prompts the user to discuss the findings with his / her healthcare provider.

[0284] 295. EXAMPLE 6:

[0285] 296. The user wishes to examine whether he / she has symptoms compatible with inflammatory bowel disease (IBD), and can do so by performing the following steps:

[0286] 297. 1. The user turns the breathalyzer device on by pressing on the power button.

[0287] 298. 2. The device gives a greeting and asks the user to select (e.g., tap) the drop-down menu.

[0288] 299. 3. From the menu the user selects IBD.

[0289] 300. 4. The device responds with the following process:

[0290] 301. A) Asks for information and whether the user has used antibiotics or any other medications which treat IBD;

[0291] 302. B) Depending on the answer, the device responds with the process.

[0292] 303. 5. The user returns after overnight fast (8hrs.) in the morning after brushing teeth but not using mouthwash.

[0293] 304. 6. The user turns the device on.

[0294] 305. 7. The user selects (e.g., taps) the menu.

[0295] 306. 8. The user selects (e.g., taps) IBD.

[0296] 307. 9. The device asks if the user is ready.

[0297] 308. 10. The user responds yes.

[0298] 309. 11. The device asks the user to exhale for 5 seconds and wait.

[0299] 310. 12. The user exhales for 5 seconds.

[0300] 311. 13. The device blocks all sensors except those which are programmed by the software to operate under the IBD request. 312. 14. The device responds with the measurement of each of these gases and an assessment of whether they are within normal limits. The device prompts the user to discuss the findings with his / her healthcare provider.

[0301] 313. EXAMPLE 7:

[0302] 314. The user suffers from asthma with an inflammatory component and is required to monitor the efficacy of the treatment and wishes to prevent flare-ups of the disease. In this case, the user seeks to measure nitric oxide in his / her breath, and can do so by performing the following steps:

[0303] 315. 1. The user turns on the breathalyzer device 100 by pressing on the power button.

[0304] 316. 2. The device provides a greeting and asks the user to select (e.g., tap) the drop-down menu icon.

[0305] 317. 3. On the menu, the user selects (e.g., taps) asthma / Nitric Oxide.

[0306] 318. 4. The device returns with instructions as to how to proceed with the breath test, in this case to breathe for about 5-10 seconds into the device through its mouth piece or opening and asks the user if he or she is ready.

[0307] 319. 5. The user selects (e.g., taps) yes.

[0308] 320. 6. The device asks for input of data such as name, age, gender, height and weight.

[0309] 321. 7. After the input, the device blocks all sensors except for the one which detects nitric oxide.

[0310] 322. 8. The device measures the nitric oxide in the user’s breath sample.

[0311] 323. 9. The device stores the result and compares it to previous results from the same user.

[0312] 324. 10. The device outputs the result on the screen and gives the trendline from the previous results from tests of the past 1-365 days and provides an assessment indicating whether the results are within normal limits or not.

[0313] 325. 11. The device prompts the user to discuss the findings with his / her health care provider. 326. While some preferred embodiments of the invention have been described, it should be understood that various changes, adaptations and modifications may be made therein without departing from the spirit of the invention and the scope of the appended claims.

[0314] 327. EXAMPLE S:

[0315] 328. The user wishes to measure ammonia in the breath as a biomarker for early metabolic dysfunction associated steatohepatitis (MASH).

[0316] 329. The user turns on the breathalyzer (fig 1) which detects and measures ammonia in breath; or the user turns on the breathalyzer (fig. 21) and from the drop down menu selects to press the icon that says MASH.

[0317] 330. The user abstains from high protein for the 24 hours prior to taking the breath test using either breathalyzer (fig.1 or fig 21).

[0318] 331. After overnight fasting (nothing to eat or drink except for water) the user brushes the teeth 15 minutes prior to taking the breath test but does not use mouthwash.

[0319] 332. After waiting for 15 minutes after brushing teeth, the user turns on the breathalyzer and reads the prompt on the screen display.

[0320] 333. The user places a mouthpiece in the receptacle of breathalyzer (fig. 1) or uses the mouthpiece for breathalyzer fig. 21.

[0321] 334. The user exhales into the mouthpiece for about 5 seconds and then waits as the breathalyzer measures the ammonia in the user’s breath.

[0322] 335. The breathalyzer ends the test by providing the results and indicates whether the user has an abnormality with the ammonia in the breath.

[0323] 336. The breathalyzer, through the microprocessor and machine learning algorithms filed within the microprocessor (figs 31-34), calculates and predicts whether the user has evidence of MASH.

[0324] 337. The user is prompted to discuss the results with the health care provider.

[0325] 338. EXAMPLE 9 339. The user wishes to be tested for infection caused by C. Difficile (C.Diff. ).

[0326] 340. The user turns on the breathalyze device which is dedicated to test for a pattern of breath gases known to be elevated in C. Diff infection.

[0327] 341. The user receives instructions as to the method for breath testing for C. Diff.

[0328] 342. The user is advised to fast, except of water and necessary medications, overnight (8-12 hours). In the morning, the user brushes teeth 15 minutes before taking the test and does not use mouthwash

[0329] 343. The user exhales into the device through the removable mouthpiece once.

[0330] 344. The sensors react to the breath gases to which they are selective.

[0331] 345. The device collects, analyzes and stores the data, calculates the concentration of gas or gases and displays the result.

[0332] 346. The device prompts the user to seek medical advice if the device has detected the presence of C. Diff. in the user.

Claims

1. CLAIMSWhat is claimed:

1. An autonomous, standalone, hand-held, battery-operated breath analyzer device, which accepts exhaled breath through a removable and replaceable mouthpiece and comprises: one or more than one sensor; wherein the sensor or more than one sensor contacts the exhaled breath; wherein the sensor or more than one sensor comprises a chemical material and a conductive material; wherein the chemical material is a polymer doped with a dopant; wherein the doped polymer is selective gas material that has a resistivity that increases in response to the increased concentration of gas in the exhaled breath sample that contacts the sensor; wherein the chemical material contacts the conductive material; a microprocessor; wherein recalled data pass through a mathematical model which is coded in the microcontroller (fitting) and yield multiple key parameters; wherein the key parameters along with the user's input information pass through a trained Machine Learning model or other classification model which includes generated patients’ data and data implanted in the microcontroller; and wherein the microprocessor returns the classification result; an electrical circuit; wherein the electrical circuit operably connects the sensor to the microprocessor; wherein the microprocessor detects resistivity in the electrical circuit and uses the resistivity to calculate concertation of gas or gases in the exhaled breath sample;2. The breathalyzer of claim 1 wherein the sensor has a resistivity that increases with the increased concentration of hydrogen in the breath sample;3. The breathalyzer of claim 1 wherein the sensor has a resistivity that increases with the increased concentration of ammonia in the breath sample;4. The breathalyzer of claim 1 wherein the sensor has a resistivity that increases with the increased concentration of CO2 and13CC>2 in the breath sample;5. The breathalyzer of claim 1 wherein the sensor has a resistivity that increases with the increased concentration of methane in the breath sample;6. The breathalyzer of claim 1 wherein the sensor has a resistivity that increases with the increased concentration of ethanol in the breath sample;7. The breathalyzer of claim 1 wherein the sensor has a resistivity that increases with the increased concentration of methanol in the breath sample;8. The breathalyzer of claim 1 wherein the sensor has a resistivity that increases with the increased concentration of isoprene in the breath sample;9. The breathalyzer of claim 1 wherein the sensor has a resistivity that increases with the increased concentration of acetone in the breath sample;10. The breathalyzer of claim 1 wherein the sensor has a resistivity that increases with the increased concentration of oxygen in the breath sample;11. The breathalyzer of claim 1 wherein the sensor has a resistivity that increases with the increased concentration of nitrogen in the breath sample;12. The breathalyzer of claim 1 wherein the sensor has a resistivity that increases with the increased concentration of ethanol in the breath sample;13. The breathalyzer of claim 1 wherein the sensor has a resistivity that increases with the increased concentration of nitric oxide in the breath sample;14. The breathalyzer of claim 1 wherein the polymer is polyaniline;15. The breathalyzer of claim 1 wherein the polymer is polypyrolle;16. The breathalyzer of claim 1 wherein the dopant is an acid;17. The breathalyzer of claim 16 wherein the dopant acid is Camphorsulfonic acid (CSA) or Dinonylnapthalenesulfonic (DNNSA) acid or dodecylbenzynesulfonic (DBSA) acid or hydrochloric acid (HCL) or another type of acid;18. The breathalyzer of claim 14 wherein polyaniline is doped with CSA and re-doped with HCL19. The breathalyzer of claim 1 wherein the sensor has a resistivity that increases with the increase of acetone in the breath sample;20. The breathalyzer of claim 1 wherein the replaceable mouthpiece is securely attached to the body of the breathalyzer;21. The breathalyzer of claim 1 wherein the sensors have a resistivity that increases with the increase of acetaldehyde, 2-propanol, acetonitrile, acrylonitrile, benzene, isoprene, pentane, methylexane, ethane, hydrogen sulfide, triethyl amine, trimethylamine, carbon disulfide, dimethyl sulfide, 1 -Heptene, 1 -Octene, 1 -Nonene, 1 -Decene, octane, nonene, dodecane, cyclohexane, 2-butane, indole, ester, carbon disulfide, pentane, nitric oxide (NO), ethane and propane in the breath sample22. The breathalyzer of claim 1 wherein the polymer is doped polyaniline;23. The breathalyzer of claim 1 wherein the polymer is doped polypyrrole;24. The breathalyzer of claim 1 wherein the polymer is sulfonated polyaniline;25. The breathalyzer of claim 1 wherein the conductive material comprises a plurality of electrodes;26. The breathalyzer of claim 25 wherein the plurality of electrodes is finger electrodes separated by distance between them27. The breathalyzer of claim 1, wherein the trained file catalogs the exhaled breath that contacts the resistivity of the sensor that contacts the exhaled breath;28. The breathalyzer of claim 27, wherein the microprocessor and the integrated circuit predict the presence of disease in the person whose exhaled breath contacts the sensor.

29. An autonomous, standalone, hand-held, battery-operated breath analyzer device, which accepts exhaled breath through a removable and replaceable mouthpiece and comprises: one or more than one sensor; wherein the sensor or more than one sensor contacts the exhaled breath; wherein the sensor or more than one sensor comprises a chemical material and a conductive material; wherein the chemical material is a polymer doped with a dopant; wherein the doped polymer is selective gas material that has a resistivity that increases in response to the increased concentration of gas in the exhaled breath sample that contacts the sensor; wherein the chemical material contacts the conductive material; a microprocessor; wherein recalled data pass through a mathematical model which is coded in the microcontroller (fitting) and yield multiple key parameters; wherein the key parameters along with the user's input information pass through a trained Machine Learning model or other classification model which includes generated patients’ data and data implanted in the microcontroller; and wherein the microprocessor returns the classification result; an electrical circuit; wherein the electrical circuit operably connects the sensor to the microprocessor; wherein the microprocessor detects resistivity in the electrical circuit and uses the resistivity to calculate concertation of gas or gases in the exhaled breath sample;30. The breathalyzer of claim 27 wherein the one or more than one sensors include sensors that have resistivity that increases with the increase of concentration of each of the following gases: 1 -iodononane and benzene, Nonanal, 4-Methyl -2 -pentanone, Decanal, 2 -Methylbutane, 1,2-Pentadiene, 2-Methylpentane, 3 -Methylpentane, Methylcyclopentane, Cyclohexane, Methylcyclohexane, 1,3-Dimethylbenzene, 4-Methyloctane, 1,4-Dimethylbenzene, 4- methylundecane, trimethyldecane.

31. The breathalyzer of claim 30 wherein the user is tested for colorectal cancer;32. An autonomous, standalone, hand-held, battery-operated breath analyzer device, which accepts exhaled breath through a removable and replaceable mouthpiece and comprises: one or more than one sensor; wherein the sensor or more than one sensor contacts the exhaled breath; wherein the sensor or more than one sensor comprises a chemical material and a conductive material; wherein the chemical material is a polymer doped with a dopant; wherein the doped polymer is selective gas material that has a resistivity that increases in response to the increased concentration of gas in the exhaled breath sample that contacts the sensor; wherein the chemical material contacts the conductive material; a microprocessor; wherein recalled data pass through a mathematical model which is coded in the microcontroller (fitting) and yield multiple key parameters; wherein the key parameters along with the user's input information pass through a trained Machine Learning model or other classification model which includes generated patients’ data and data implanted in the microcontroller; and wherein the microprocessor returns the classification result; an electrical circuit; wherein the electrical circuit operably connects the sensor to the microprocessor; wherein the microprocessor detects resistivity in the electrical circuit and uses the resistivity to calculate concertation of gas or gases in the exhaled breath sample;33. The breathalyzer of claim 32 wherein the one or more than one sensor include sensors that have resistivity in response to the increased concentration of each of the following gases: propan- l-ol, 3 -methylbutanal, ethyl propionate, hexanoic acid, 4-methyl phenol, dodecane, and indole, methanol, p-cresol, dimethylamine and a range sulfur compounds (ethylene sulfide, dimethylsulfide and methyl thioacetate).

34. The breathalyzer of claim 33 wherein the user is tested for C. Difficile infection.

35. An autonomous, standalone, hand-held, battery-operated breath analyzer device, which accepts exhaled breath through a removable and replaceable mouthpiece and comprises: one or more than one sensor; wherein the sensor or more than one sensor contacts the exhaled breath; wherein the sensor or more than one sensor comprises a chemical material and aconductive material; wherein the chemical material is a polymer doped with a dopant; wherein the doped polymer is selective gas material that has a resistivity that increases in response to the increased concentration of gas in the exhaled breath sample that contacts the sensor; wherein the chemical material contacts the conductive material; a microprocessor; wherein recalled data pass through a mathematical model which is coded in the microcontroller (fitting) and yield multiple key parameters; wherein the key parameters along with the user's input information pass through a trained Machine Learning model or other classification model which includes generated patients’ data and data implanted in the microcontroller; and wherein the microprocessor returns the classification result; an electrical circuit; wherein the electrical circuit operably connects the sensor to the microprocessor; wherein the microprocessor detects resistivity in the electrical circuit and uses the resistivity to calculate concertation of gas or gases in the exhaled breath sample;36. The breathalyzer of claim 35 wherein the one or more than one sensor includes sensors that have a resistivity that increases as a result of increased concentration of each of the following gases: Nitrogen, oxygen, CO2, methane, acetone, isoprene methanol and ethanol.

37. The breathalyzer of claim 36 wherein the performance of an athlete is predicted on the basis of the pattern of each of the measured gases.

38. Abreath test method for predicting athletic performance comprising steps of: a. Providing an autonomous breath testing device that includes a removable and replaceable mouthpiece and a main body, wherein the main body includes one or more than one sensor, software enhanced with machine learning algorithms and a processor and an electrical circuit. Wherein the one or more than one sensor comprise gas- selective material that has a resistivity that increases as a result of the increase in concentration of breath gas to which it is selective; b. Prompting the user to exhale into the breath testing device before the beginning of exercise, during exercise and after exercise is completed at 10 minutes, 30 minutes and 60 minutes; c. Allowing the processor to measure resistivity of the one or more than one sensor that occurs when the breath contacts the one or more than one sensors; d. Display the results;e. Transmit the data to a smart phone, computer or the Cloud.

39. Awearable, autonomous, breath testing device that comprises: a. one or more than one gas-selective sensors wherein the sensors have a resistivity that increases as a result of the increased concentration of the breath gas or gases; b. Software enhanced with machine learning algorithms; c. A processor that detects resistivity; and d. Electrical circuit that operably connects the sensors to the processor.

40. The breath testing device of claim 39 wherein the device comprises of one or more than layers of face covering;41. The breath testing device of claim 40 wherein the face covering includes two layers;42. The breath testing device of claim 41 wherein one layer includes the sensors, desiccant and connectors and the second layer includes the software, processor and electrical circuit;43. The breath testing device of claim 40 wherein the face covering covers the mouth and the nose;44. The breath testing device of claim 40 wherein the face covering covers the mouth only;45. The breath testing device of claim 40 wherein the face covering covers the nose only.

46. Awearable autonomous breath testing device that comprises of: a. A removable and replaceable mouthpiece; and b. A main body that includes the on / off switch, the opening for the mouthpiece and he mouthpiece cover, one or more than one sensor, software with machine learning enhanced algorithms and processor, and an electrical circuit that operably connects the one or more than one sensor to the processor and a screen; wherein the one or more than one sensor has a resistivity that increases with the increase of concentration of the breath gas or gases;47. The breath testing device of claim 46 wherein the device is worn on part of the body;48. The breath testing device of claim 47 wherein the part of the body is the wrist;49. The breath testing device of claim 46 wherein the screen is LED screen;50. A breath test method for screening for a health disorder or athletic or exercise performance that comprises of the following steps: a. Providing an autonomous wearable breath testing device that comprises of: i. A removable and replaceable mouthpiece; andii. A main body that includes the on / off switch, the opening for the mouthpiece and the mouthpiece cover, one or more than one sensor, software with machine learning enhanced algorithms and processor, and an electrical circuit that operably connects the one or more than one sensor to the processor; wherein the one or more than one sensor has a resistivity that increases with the increase of concentration of the breath gas or gases; b. Prompting the user to turn on the wearable device through the on / off switch; c. Prompting a user to remove the mouthpiece cover and insert the mouthpiece firmly into the mouthpiece opening; d. Prompting the user to exhale into the mouthpiece; e. Allowing the processor to measure resistivity from the sensor or sensors that came in contact with the breath; f. Allowing the device to collect the data, store the data, calculate the result, display the result and wirelessly transmit the result to smart phone, computer or Cloud;51. The method of claim 50 wherein the health disorder is gastrointestinal disorder;52. The method of claim 50 wherein the health disorder is a liver disorder;53. The method of claim 50 wherein the health disorder is renal disorder;54. The method of claim 50 wherein the health disorder is metabolic disorder;55. The method of claim 50 wherein the health disorder is infection;56. The method of claim 50 wherein the health disorder is cancer57. The method of Claim 50 wherein the exercise is running;58. The method of Claim 50 wherein the exercise is basketball;59. The method of Claim 50 wherein the exercise is baseball;60. The method of Claim 50 wherein the exercise is playing tennis;61. The method of Claim 50 wherein the exercise is soccer;62. The method of Claim 50 wherein the exercise is football;63. The method of Claim 50 wherein the exercise is gymnastics;64. The method of Claim 50 wherein the exercise is general exercise e.g., walking65. The method of Claim 50 wherein the exercise is dancing;66. The method of Claim 65 wherein the exercise is aerobic dancing.

Images