Exhalation detection device and analysis

Abstract

This invention provides an improved breath test instrument and method for determining the presence of diseases in the human body, including but not limited to detecting Helicobacter pylori infection, celiac disease, metabolic disorder-associated steatohepatitis (MASH), and inflammatory bowel disease (IBD) in the digestive tract of a subject. In some embodiments, this invention provides a universal breath test platform and method for detecting diseases of the gastrointestinal tract, liver, kidneys, and lungs, as well as cancer, infectious diseases, and metabolic diseases.

Description

Cross-references to related applications

[0001] This application claims the benefit of U.S. Provisional Application Serial No. 63 / 559,446, filed on February 29, 2024, which is incorporated herein by reference in its entirety. Technical Field

[0002] This application generally relates to a breath testing device and method for detecting various diseases of the gastrointestinal tract, liver, lungs, and kidneys, as well as cancer, infectious diseases, and metabolic diseases. This application also relates to a breath testing instrument and method for detecting various gases in a human breath sample to determine the presence of diseases in the digestive tract of a subject. In some instances, diseases may include, but are not limited to, Helicobacter pylori, celiac disease, metabolic disorder-associated steatohepatitis (MAHD), and inflammatory bowel disease (IBD). Background Technology

[0003] Exhaled breath contains numerous gases and thousands of volatile organic compounds (VOCs). These gases and VOCs can be detected using techniques such as gas chromatography-mass spectrometry (GC-MS), selective ion flow tube mass spectrometry (SIFT-MS), FTIR spectroscopy, ion mobility mass spectrometry, field asymmetric ion mobility spectrometry, semiconductor chips, carbon nanotubes, metal oxides, doped and undoped polymers, and other types of conductive materials sensitive to various gases. Types of conductive materials include chemically sensitive field-effect transistors (CFETs), floating-gate CFETs, or any other field-effect transistor. Other types of gas sensors are optical, electrochemical, thermochemical, and surface acoustic wave (SAW) films deposited on conductive materials such as gold, platinum, palladium, or other metals.

[0004] Exhaled breath primarily consists of oxygen (O2) 16%, carbon dioxide (CO2) 4%, nitrogen (N2) 75%, and water vapor (5%-6%). In addition, exhaled breath contains trace amounts of argon, hydrogen (H2), ammonia (NH3), acetone, methanol, ethanol, and methane (CH4), which are some of the most commonly encountered volatile organic compounds (VOCs). Other VOCs in exhaled breath include acetaldehyde, 2-propanol, acetonitrile, acrylonitrile, benzene, isoprene, pentane, methylhexane, ethane, hydrogen sulfide, triethylamine, trimethylamine, carbon disulfide, dimethyl sulfide, 1-heptene, 1-octene, 1-nonene, 1-decene, octane, nonene, dodecane, cyclohexane, 2-butene, indole, esters, carbon disulfide, pentane, nitric oxide (NO), ethane, and propane. Exhaled VOCs are metabolic byproducts of physiological or pathophysiological processes occurring in the digestive tract, small intestine, and large intestine in healthy or diseased individuals.

[0005] Metabolomics is the field of research on byproducts or metabolites of physiological or pathophysiological processes in the human body. These byproducts are volatile organic compounds (VOCs), which originate from the breakdown of substances ingested by bacteria in the colon of healthy individuals and those suffering from gastrointestinal, kidney, lung, metabolic, or liver diseases, as well as metabolic syndrome, obesity, cancer, and infections. Analysis of VOCs and their spectra reflects the composition of the microbiome and is considered a biomarker for these diseases. Currently, VOC detection is performed using techniques such as gas chromatography-mass spectrometry (GC-MS), selective ion flow tube mass spectrometry (SIFT-MS), field asymmetric ion mobility spectrometry (FAIMS), and electronic noses (e-Nose).

[0006] Technologies such as GC-MS, SIFT-MS, and FAIMS are effective at detecting VOCs, especially those derived from the colonic microbiota. However, these instruments are expensive, bulky, and mostly stationary. Electronic nose technology (a pattern recognition technology) uses a variety of sensors with low gas selectivity and instability. None of these technologies use a separate sensor to detect every VOC in exhaled gas, and these instruments are neither small, handheld, nor inexpensive, nor can they be used over-the-counter (OTC) by exhaling directly through the instrument's opening or mouthpiece. Metal oxide sensors are not optimal, either alone or in combination with GC-MS, as most of them do not operate at room temperature, and most require heating to approximately 450 degrees Celsius before use.

[0007] Celiac disease is an autoimmune enteropathy triggered by the intake of gluten (particularly its immunotoxic component, prolycidin), primarily affecting susceptible individuals. Celiac enteropathy can be managed with a complete and lifelong gluten-free diet. Failure to adhere to or strictly follow a gluten-free diet may lead to small intestinal lymphoma, adenocarcinoma, or other autoimmune diseases such as diabetes, thyroiditis, and hepatitis. Individuals susceptible to celiac disease express autoantibodies such as anti-endothelial antibodies and anti-tissue transglutaminase. In addition to those diagnosed with celiac disease, there are subclinical or asymptomatic celiac patients, as well as family members of celiac patients who, while asymptomatic, exhibit histological abnormalities in the small intestine and remain undiagnosed. Undiagnosed patients and those with delayed diagnosis continue to consume gluten-containing diets and are at risk of severe damage to their intestinal mucosa and life-threatening consequences such as cancer.

[0008] The incidence of celiac disease is rising in regions worldwide where gluten consumption is widespread. Currently, the prevalence of celiac disease in the United States is 1%, significantly higher than the 0.02% in the early 1990s. The incidence of non-celiac gluten sensitivity (NCGS) is also rising, estimated at 3%–6% in the United States and Europe. NCGS patients do not have autoantibodies or intestinal mucosal changes characteristic of celiac disease, but they experience symptoms similar to those of celiac disease after consuming gluten, and their symptoms improve after adopting a gluten-free diet.

[0009] The current gold standard for diagnosing celiac disease is a small bowel biopsy obtained via upper gastrointestinal endoscopy. Patients with celiac disease who must adhere to a strict lifelong gluten-free diet undergo follow-up endoscopy combined with biopsy to assess their adherence to the gluten-free diet and the health of the small bowel mucosa. Other diagnostic methods exist, including blood tests for anti-endothelial antibodies and anti-transglutaminase antibodies. Currently, there are no screening tests for patients with symptomatic or asymptomatic celiac disease (i.e., subclinical celiac disease and asymptomatic family members), for whom there is no reason to undergo endoscopy. However, these asymptomatic patients have a high risk of complications similar to those in symptomatic celiac disease and may develop cancer. These asymptomatic patients, who are mostly family members of patients with celiac disease, can benefit from certain embodiments of the present invention, which provides a non-invasive breath test with high sensitivity for hydrogen in exhaled breath after an eight- to twelve-hour (8-12)-hour overnight fast.

[0010] Helicobacter pylori (H. pylori), affecting two-thirds of the world's population, is a highly contagious Gram-negative bacterium that causes chronic gastritis, peptic ulcers, and may lead to gastric cancer and other malignancies such as gastric lymphoma (MALT lymphoma). This bacterium is also associated with extraintestinal diseases such as anemia, liver disease, gallbladder disease, and lung diseases such as asthma. One characteristic of H. pylori is its ability to hydrolyze urea into CO2 and ammonia (NH3) using its abundant urease, as shown in the following equation: CO(NH2)2 + HOH → urease → CO2 + 2NH3.

[0011] The gold standard for diagnosing Helicobacter pylori is 13 The urea breath test (UBT) is a test performed after urea ingestion. 13 After measuring the exhaled urea with C-labeled urea... 13 CO2, the reaction equation is as follows: The current 13CUBT requires relatively expensive reagent kits and costly equipment (thousands of dollars) that must be operated by professionals. Other known methods for diagnosing Helicobacter pylori infection include: 1) upper gastrointestinal endoscopy combined with biopsy and tissue culture; 2) upper gastrointestinal endoscopy combined with biopsy and rapid urease test (CLO test); and 3) serum antibodies against Helicobacter pylori. Positive antibodies indicate Helicobacter pylori infection. However, antibodies may remain positive even after the bacteria are eradicated. Therefore, serum antibodies cannot be compared to... 13 C UBT (and other methods) are used to confirm eradication. Existing diagnostic methods are not inexpensive and are not handheld, meaning the test and device cannot be integrated into one unit.

[0012] Inflammatory bowel diseases primarily include Crohn's disease and ulcerative colitis. Each of these inflammatory bowel diseases has different characteristics associated with the following: 1) different locations of the intestine affected by inflammation (ulcerative colitis is limited to the large intestine, while Crohn's disease may involve the entire digestive tract, but mainly the small intestine); 2) different histological changes in the intestinal mucosa; 3) different diagnoses; 4) different treatments; and 5) different prognoses.

[0013] The diagnosis of inflammatory bowel disease (IBD) and the differentiation between Crohn's disease and ulcerative colitis are determined through endoscopy of the small and large intestines, combined with biopsy and histological examination of the intestinal mucosa, as well as radiological studies (especially in Crohn's disease). Once diagnosed, appropriate treatment is initiated. This treatment is lengthy and requires regular follow-up via endoscopy and biopsy to assess its effectiveness.

[0014] Alterations in the colonic metabolome have been observed in patients with inflammatory bowel diseases (Crohn's disease and ulcerative colitis), contributing to the formation of a volatile organic compound (VOC) profile in their exhaled breath. These VOCs include acetaldehyde, 2-propanol, acetonitrile, acrylonitrile, benzene, isoprene, pentane, methylhexane, ethane, hydrogen sulfide, triethylamine, trimethylamine, carbon disulfide, dimethyl sulfide, 1-heptene, 1-octene, 1-nonene, and 1-decene, among others.

[0015] Small intestinal bacterial overgrowth (SIBO) occurs in susceptible individuals and presents with symptoms similar to inflammatory bowel disease, such as abdominal pain, diarrhea, anemia, and weight loss. The small intestine typically contains a very small number of microorganisms compared to the large intestine, which usually contains trillions (estimated at 100 billion per milliliter). SIBO occurs when bacteria that normally ferment carbohydrates in the large intestine migrate to the small intestine. As a result, carbohydrate fermentation occurs in the small intestine, which in turn causes symptoms such as abdominal pain, steatorrhea, and abnormally high hydrogen production, which is absorbed and released through the lungs during respiration.

[0016] Conditions that can lead to SIBO include anatomical abnormalities, abnormal intestinal motility (such as pseudo-obstruction), absence of the migration motor complex, autonomic neuropathy (such as diabetes), excessive bacterial load (such as achlorhydria), fistulas and ileocecal valve atresia, and immune system problems (such as immunodeficiency and malnutrition).

[0017] The diagnosis of SIBO is obtained through invasive or non-invasive examinations. Invasive examinations include small bowel endoscopy and aspiration culture of contents to detect aerobic and anaerobic bacteria and to perform kinematic studies. Non-invasive examinations include 72-hour fecal collection of fecal fat, serum bile acids, and a breath test. The most common breath test is the hydrogen breath test performed after lactulose ingestion. Lactulose (a non-absorbable sugar) provides a substrate for colonic bacteria to produce hydrogen, leading to an increase in hydrogen concentration in the breath. If E. coli are present in the small intestine, hydrogen production occurs much earlier than normal after lactulose ingestion, and early and late peaks of hydrogen production are observed in the breath test. The gold standard hydrogen breath test is performed using a Quintron breath analyzer. Individuals undergoing the SIBO breath test provide a fasting baseline breath sample and ingest a certain amount (approximately 7 g / kg) of lactulose. In the case of SIBO, hydrogen gas produces two peaks after lactulose excretion: one earlier because coliform bacteria are present in the small intestine; and the other later, when lactulose reaches the large intestine.

[0018] Irritable bowel syndrome (IBS) is a diagnosis of exclusion and is applicable to individuals presenting with symptoms of abdominal pain, diarrhea or constipation, bloating, and flatulence. IBS is a functional disorder of the digestive tract that, although its symptoms are similar to those of IBD, has no organic cause and no histological abnormalities of the digestive tract are found. Abnormal bowel motility and food intolerances have been observed in some individuals with IBS, which may manifest as diarrhea or constipation. The diagnosis of IBS can only be made after ruling out all organic causes of IBD, SIBO, infection, and celiac disease. Analysis of fecal volatile organic metabolites has been reported to have a spectrum consistent with diarrheal IBS (IBS-D), distinguishing IBS-D individuals from IBD individuals and healthy individuals, with a sensitivity of 94%–96% and a specificity of 80%–82%. IBS-D fecal volatile organic compounds contain short-chain fatty acid esters, such as cyclohexanecarboxylic acid, butyrate, acetate, and propionate. This observation could facilitate the development of colon-derived expiratory metabolomics as a biomarker for IBS, particularly diarrheal IBS.

[0019] Lactose intolerance is the intolerance to lactose, a carbohydrate found in milk. Symptoms of lactose intolerance are similar to those of irritable bowel syndrome (IBS) and other malabsorption problems. These symptoms include abdominal pain, diarrhea, and bloating, and occur after consuming milk or dairy products. Lactose is a disaccharide that is broken down into the monosaccharides glucose and galactose by lactase, an enzyme present at the brush border of the intestine. Glucose and galactose are readily absorbed and usually do not cause symptoms. In individuals with a deficiency or severe deficiency of lactase, such as in hereditary conditions or after infections involving the digestive tract (such as viral enteritis or parasitic or microbial infections), lactose cannot be metabolized and leads to symptoms similar to those of carbohydrate malabsorption, such as diarrhea, abdominal pain, bloating, and flatulence. Fructose intolerance is the inability to tolerate the intake of fructose, a carbohydrate found in fruits. Therefore, individuals with this disorder experience symptoms similar to those of individuals with lactose intolerance after consuming fruit. It is estimated that the adult gut can tolerate up to 25 g of fructose without symptoms. Ingesting more than 25 g of fructose may cause diarrhea, bloating, and abdominal pain. Any amount of fructose can trigger symptoms in individuals with hereditary fructose intolerance due to a deficiency in the Aldolase B enzyme gene. Liver diseases are divided into two categories: hereditary and congenital liver diseases, and acquired liver diseases. Hereditary and congenital liver diseases include carbohydrate metabolism disorders, amino acid metabolism disorders, mitochondrial fatty acid β-oxidation abnormalities, α1-antitrypsin deficiency, bile acid synthesis and metabolism disorders, Wilson's disease, etc. Acquired liver diseases are mainly caused by viral infections, such as hepatitis A, B, C, and other viruses, which can lead to chronic hepatitis, cirrhosis, and end-stage liver disease, hepatocellular carcinoma (HCC) and other liver malignancies, as well as liver diseases caused by obesity, such as metabolically associative fatty liver disease (MASLD), also known as NAFLD, and metabolically associative fatty liver disease (MASH), also known as NASH, and metabolic abnormalities such as diabetes. Autoimmune hepatitis, which can lead to chronic hepatitis and cirrhosis, is an inflammatory liver disease secondary to an autoimmune disease affecting more than one organ in the body. HCC can be caused by viral hepatitis or steatohepatitis, which is due to fat deposition in the tissues. Steatohepatitis may be secondary to obesity, diabetes, and alcoholic hepatitis. If left untreated, steatohepatitis can lead to cirrhosis and end-stage liver disease. Metabolic changes have been observed in hereditary liver diseases, as well as acquired and end-stage liver diseases. Encephalopathy caused by hyperammonemia is one of the symptoms of liver failure.

[0020] Kidney disease refers to diseases of the kidneys that can be hereditary, idiopathic, or acquired. Hereditary kidney diseases include several types of glomerulonephritis, such as IgA nephropathy, Alport syndrome, pulmonary hemorrhage-nephritis syndrome, membranous glomerulonephritis, and lupus-associated glomerulonephritis, as well as diseases involving abnormal kidney anatomy, such as polycystic kidney disease and UPJ obstruction. Acquired kidney diseases include post-infectious glomerulonephritis, such as acute post-streptococcal glomerulonephritis, hemolytic uremic syndrome (HUS), drug-induced glomerulonephritis, and post-traumatic nephropathy. Symptoms of kidney disease include hematuria, proteinuria, edema, and growth retardation in children. When kidney disease becomes chronic and progresses to kidney failure (renal failure), dialysis and eventually kidney transplantation become necessary. Hyperammonemia is a key characteristic of end-stage renal failure because the kidneys are unable to excrete accumulated ammonia.

[0021] Human exhalation comprises over 3,500 components (gases), most of which are volatile organic compounds (VOCs) in trace amounts. Besides nitric oxide (NO), which has become a biomarker for respiratory inflammation and asthma, a VOC profile is also prevalent in asthma populations with both allergic and inflammatory components. These compounds include nonane, 2,2,4,6,6-pentamethylheptane, decane, 3,6-dimethyldecane, dodecane, and tetradecane. The expiratory metabolome provides characterization for chronic obstructive pulmonary disease (COPD).

[0022] Metabolic diseases such as diabetes and obesity affect the metabolome. Elevated acetone levels in the breath of individuals with diabetes are consistent with undiagnosed diabetes or poorly managed diabetes in diagnosed individuals. The concentration of acetone in the breath has been found to correlate with the concentration of β-hydroxybutyrate in the venous blood of obese patients in a fasting state. Studies have identified eight specific metabolites, such as isopropanol and 2,3,4-trimethylhexane, 2,6,8-trimethyldecane, tridecane, and undecane, which provide a more specific spectrum for diagnosing the presence of type 2 diabetes, exhibiting both high sensitivity and specificity. Elevated VOCs have been described in patients with various cancers, such as colon cancer, lung cancer, and breast cancer. Infections involving the gastrointestinal tract, such as bacterial infections (Salmonella, Shigella) and parasitic infections, alter colonic bacteria and lead to changes in the corresponding expiratory metabolome.

[0023] Helicobacter pylori (“H. pylori”) is a Gram-negative bacterium found in the digestive tract, affecting approximately two-thirds of the world’s population. Most people are infected with H. pylori in childhood and may never experience any signs or symptoms of infection. However, when signs or symptoms of H. pylori infection do appear, they may include burning pain in the abdomen and chest, nausea, vomiting, frequent hiccups, bloating, and weight loss. H. pylori causes gastritis and most peptic ulcers and is associated with other intestinal diseases such as inflammatory bowel disease, hepatobiliary diseases, and pancreatic diseases. H. pylori is also associated with extraintestinal diseases, some of which are idiopathic thrombocytopenic purpura, iron deficiency anemia, kidney disease, and cardiovascular diseases such as ischemic heart disease and atherosclerosis.

[0024] Furthermore, Helicobacter pylori was classified as a Group 1 carcinogen for humans by the International Agency for Research on Cancer (IARC) in 1994 and is closely associated with the development of gastric cancer and gastric MALT lymphoma (a type of lymphoma). Helicobacter pylori is also associated with other cancers such as colon cancer and pancreatic cancer.

[0025] Current diagnostic methods for detecting Helicobacter pylori infection have several shortcomings. Some existing diagnostic methods include endoscopic biopsy and bacterial culture. These methods are expensive, cumbersome, and pose certain risks to the subject. Furthermore, results are typically available several days after the test. In addition, these invasive methods utilize large and expensive equipment, are performed by highly trained personnel, require anesthesia, sedation, or both, and necessitate the use of pathology and microbiology laboratories for diagnosis.

[0026] Currently available non-invasive methods for detecting Helicobacter pylori include: 13 C-labeled urea breath test, detection of Helicobacter pylori antigen in stool, detection of Helicobacter pylori DNA in stool, and serum antibody detection are all known non-invasive methods. These methods are costly, cannot be self-tested, and results typically take hours or even days to obtain. Currently, these non-invasive methods utilize laboratory equipment to process breath samples... 13 CO2 analysis, blood tests for Helicobacter pylori antibodies, and stool tests for Helicobacter pylori antigens and / or Helicobacter pylori DNA. All of these methods require expensive equipment and qualified personnel to perform the specific tests.

[0027] 13 The urea breath test is also known and has been used to detect Helicobacter pylori infection. In this known type of urea breath test, the presence of Helicobacter pylori is based on the ingestion of a urea-labeled... 13 After measuring the urea substrate of C in the subject's exhaled breath... 13 CO2: 12The ratio of CO2. Subjects exhale into a bag, which is then attached to large equipment (e.g., a spectrometer) for analysis of the exhaled breath sample. If 13 CO2: 12 A CO2 ratio above a certain value indicates that the subject is considered positive for Helicobacter pylori. However, these known types of urea breath tests have several drawbacks. First, they use labeled... 13 C's urea substrate, although 13 C is a naturally occurring isotope of carbon, but its cost is far higher than that of naturally occurring carbon in unlabeled urea. 12 C. In addition, it is known 13 The C-urea breath test requires expensive equipment (e.g., an infrared spectrometer) for analysis. 13 CO2 and 12 CO2 levels.

[0028] As described in this disclosure, it is desirable to provide a breath test method that measures ammonia and CO2 in the breath of a subject before and after the subject ingests a urea substrate. 12 CO2 or 13 CO2, 12 CO2 and 13 CO2; or both; 13 CO2: 12 The ratio of CO2 to urea levels is measured. A breath test that can be used without distinguishing between urea-labeled and urea-based substrates is also desired. Furthermore, a reliable, portable diagnostic test that can be self-administered using a handheld device to determine the presence of Helicobacter pylori is desired. Additionally, an improved breath test that is less expensive and more convenient than existing breath tests is also desired.

[0029] Breath analysis is used to detect and measure biomarkers of disease. The exhaled metabolome (which consists of a series of gases present in exhaled breath) can serve as a biomarker for several diseases of the digestive tract, liver, pancreas, kidneys, as well as cancer and infections.

[0030] One expiratory biomarker is fasting expiratory hydrogen (FBH), a diagnostic biomarker for celiac disease. Fasting expiratory hydrogen (FBH) levels are significantly elevated in patients with celiac disease compared to those with other malabsorption syndromes, and remain elevated throughout a period of strict adherence to a gluten-free diet for at least 6 months. Because FBH levels are shown to correlate with the histological condition of the small intestinal mucosa, they can be used to monitor celiac disease patients on a strict gluten-free diet. Higher FBH levels are associated with greater histological abnormalities in the small intestine; conversely, lower FBH levels are associated with less histological abnormalities. When FBH is within normal limits, the small intestinal histology is normal.

[0031] Another exhaled biomarker is fasting breath ammonia (FBA). Metabolic dysfunction-associated steatohepatitis (MASH, also known as NASH) falls under the spectrum of metabolic dysfunction-associated fatty liver disease (MASLD, also known as NAFLD). This is due to the rising global obesity rate and metabolic syndrome. 6 NAFLD has become the most common liver disease in the world. When the liver accumulates lipotoxic lipids due to fatty infiltration, it develops into MASH, leading to severe hepatocellular damage.

[0032] Lipotoxic lipids → hepatocellular damage → MASH → fibrosis → cirrhosis → HCC

[0033] Lipotoxic lipids (diacylglycerols, ceramides, etc.) mediate a series of intracellular processes (endoplasmic reticulum stress, mitochondrial dysfunction, inflammatory response, and apoptosis), leading to the histological phenotype of MASH. These intracellular processes are inducing fibrosis and may contribute to the development of hepatocellular carcinoma (HCC).

[0034] It is estimated that 2%-5% of the US population, or 20% of those with obesity-induced MASLD, will progress to MASH and cirrhosis. Furthermore, there is no clearly defined and universally accepted predictable standard. Furthermore, MASH may soon become the primary indication for liver transplantation. Without treatment, the annual cumulative risk of HCC in patients with MASH fibrosis ranges from 2.4% to 12.8%, and the estimated annual economic impact of MASLD-MASH in the United States is $103 billion. Therefore, it is urgent to treat cirrhosis before it develops. Identify and treat MASH.

[0035] Liver biopsy is an invasive procedure and the gold standard for diagnosing and staging NASH fibrosis, often used in conjunction with serum alanine aminotransferase (ALT) elevation ≥50 IU / L and imaging studies (ultrasound-MRI). Biomarkers are expected to improve the stratification of MASLD disease severity and identify additional pathways for targeted therapy before disease progression to MASH fibrosis and cirrhosis. A rigorous review of the literature reveals that several non-invasive biomarkers and combinations of biomarkers reflecting potential MASH disease pathways via blood tests are currently under development. Biomarkers such as the CK-18 fragment produced by caspase (CK-18), fibroblast growth factor 21 (FGF21), insulin-like growth factor 2 (IGF-2), and epidermal growth factor receptor (EGFR) have moderate diagnostic and prognostic accuracy. Combinations of biomarkers such as the AST:ALT ratio, MASH fibrosis score, and BARD score (BMI, AST:ALT ratio, diabetes) have lower accuracy than specific fibrosis markers (FibroTest, FibroMeter). These specific biomarkers of fibrosis can only accurately predict the stage of fibrosis after it has developed for a long time. Hyaluronic acid (HA, a major component of the extracellular matrix) was detected in the late stages of fibrosis.

[0036] Imaging biomarkers such as FibroScan and point shear wave elastography (psWE) have moderate to high accuracy in diagnosing advanced fibrosis or cirrhosis, but are less effective in diagnosing early fibrosis. Magnetic resonance elastography (MRE) has higher diagnostic success rates and accuracy than ultrasound-based techniques, but is limited by cost and availability. Genetic and genomic biomarkers are being developed to assess disease susceptibility and severity, but their accuracy and reproducibility in predicting MAFLD and MASH and their severity remain limited. Metabolomics research is underway, and studies targeting microbiome profiling have yielded an algorithm capable of predicting advanced fibrosis, suggesting that detection based on this algorithm could become a useful biomarker. However, such biomarkers cannot predict early fibrosis. Despite promising clinical studies, an accurate, reproducible, and non-invasive method for predicting early NASH has not yet been identified.

[0037] There is a need for an accurate, non-invasive, repeatable, and specific biomarker that can be used in the earliest stages of fibrosis. The stage has high predictive value for MASH fiberization.

[0038] Ammonia in MASH: Currently, no existing biomarkers are detected by breath testing, which is the simplest method for detecting potential biomarkers. Recent strong evidence from animal and human studies indicates that hepatocellular damage leads to structural changes (gene expression, activity) in the urea cycle enzymes ornithine transcarbamate (OTC) and carbamate phosphosynthase (PS), thereby hindering the conversion of ammonia to urea and causing hyperammonemia. Studies have shown a direct link between elevated ammonia concentrations and the development of fibrosis. When ammonia concentrations reach 50-300 μM / L, normally quiescent hepatic stellate cells (HSCs) are activated, proliferate rapidly, and synthesize a fibrotic matrix rich in type I collagen, becoming a key factor in the development of fibrosis, portal hypertension, and hepatocellular carcinoma (HCC). Hyperammonemia caused by hepatocellular damage is a key factor in the development of fibrosis in MASH. earliest stimuli one.

[0039] Exhaled ammonia as a biomarker for MASH: The toxicity of hyperammonemia makes ammonia monitoring a priority. However, according to the AASLD, blood ammonia measurements are considered unreliable due to inaccuracies related to blood collection methods (arterial blood collection is superior but painful and requires specialized training; venous blood often leads to inaccurate results) and potential errors during sample transport to the laboratory. Exhaled ammonia measurement is a simpler and more accurate alternative to blood ammonia, especially since lung ammonia is in equilibrium, and studies have shown a correlation between blood and exhaled ammonia. Summary of the Invention

[0040] Certain embodiments of this disclosure provide a universal electrochemical multianalyte breath analyzer for detecting gases in the breath of individuals suffering from gastrointestinal, renal, respiratory, metabolic, and inflammatory diseases, cancer, and infections. This universal electrochemical device may include a body housing multiple electrochemical sensors, electronics for supporting device operation, a power supply, a USB interface, and Bluetooth technology for wirelessly transferring data to another device or computer. The universal electrochemical device may have a display for user input and output of results, receiving the user's breath through an opening in the device body (acting as a mouthpiece) or through a removable mouthpiece.

[0041] Some other embodiments provide a breathalyzer including an input terminal, a first sensor, a second sensor, a processor, and circuitry. The input terminal receives a breath sample. The first sensor is in contact with the breath sample and comprises polyaniline and a conductive material. The polyaniline is in contact with the conductive material and is doped with a first dopant to improve the pH sensitivity of the polyaniline. The resistivity of the polyaniline increases in response to an increase in ammonia concentration. The second sensor is also in contact with the breath sample. The second sensor comprises polypyrrole and a conductive material. The polypyrrole is in contact with the conductive material and is doped with a second dopant to improve the pH sensitivity of the polypyrrole. The resistivity of the polypyrrole increases with an increase in carbon dioxide concentration. The circuitry operatively connects the first and second sensors to the processor. The processor detects the resistivity in the circuitry and uses this resistivity to calculate the total ammonia concentration and the total carbon dioxide concentration in the breath sample.

[0042] Other embodiments provide a handheld, portable breathalyzer including a removable mouthpiece and a body. The body includes a first sensor, a second sensor, a processor, and circuitry. The first sensor comprises polyaniline, and the polyaniline is doped with a first dopant to improve the pH sensitivity of the polyaniline. The resistivity of the polyaniline increases in response to an increase in ammonia concentration. The second sensor comprises polypyrrole, and the polypyrrole is doped with a second dopant to improve the pH sensitivity of the polypyrrole. The resistivity of the polypyrrole increases in response to an increase in carbon dioxide concentration. The circuitry operatively connects the first and second sensors to the processor. The processor detects the resistivity of the first sensor and uses that resistivity to calculate the ammonia concentration, and the processor detects the resistivity of the second sensor and uses that resistivity to calculate the carbon dioxide concentration.

[0043] Another embodiment provides a breathalyzer method. This breathalyzer method includes the step of providing a portable breathalyzer comprising a removable mouthpiece and a body. The body includes a first sensor, a second sensor, a processor, and circuitry. The first sensor comprises an ammonia-selective material whose resistivity increases in response to an increase in ammonia concentration. The second sensor comprises a carbon dioxide-selective material whose resistivity increases in response to an increase in carbon dioxide concentration. The circuitry operatively connects the first and second sensors to the processor, and the processor measures the resistivity of the first and second sensors. The method further includes prompting a subject to blow a baseline breath sample into the removable mouthpiece and allowing the processor to measure the resistivity of the first sensor when the baseline breath sample comes into contact with the first sensor. The method also includes prompting a subject to blow a breath sample after ingesting urea into the removable mouthpiece and allowing the processor to measure the resistivity of the first sensor when the breath sample after ingesting urea comes into contact with the first sensor. In addition, the breath detection method includes a step of comparing the resistivity measured in a baseline breath sample with the resistivity measured in a breath sample after urea ingestion.

[0044] Some other implementations provide a method for detecting the presence of Helicobacter pylori in the digestive tract of a subject. The method includes collecting a baseline breath sample from the subject and determining the total ammonia and carbon dioxide levels present in the baseline breath sample. The method also includes collecting a breath sample from the subject after ingesting urea and determining the total ammonia and carbon dioxide levels present in the urea-ingested breath sample. Furthermore, the method includes the step of indicating the presence of Helicobacter pylori in the digestive tract if the total ammonia and carbon dioxide levels in the urea-ingested breath sample exceed a predetermined value from the baseline breath sample.

[0045] Principal component analysis (PCA) is an analytical method used in medical testing to classify clinical research data. PCA can identify subjects with the same diagnosis and cluster them together based on the component inputs included in various studies.

[0046] Artificial intelligence and machine learning are being applied in medicine to better interpret disease test results. Machine learning algorithms are being used to examine clusters in data and detect anomalies with a precision previously unattainable by standard statistical methods. K-means and DBSCAN algorithms are being used to examine data and infer conclusions about whether test results are positive or negative.

[0047] In step 6 of the device operation procedure Figure 5 and Figure 34In some implementations of this method, the retrieved data is calibrated using mathematical model code programmed into the microcontroller, and the result is calculated (using this information). In step 7, after the data is calibrated and calculated, the result is displayed on an LED screen.

[0048] In step 6, the retrieved data is encoded into a mathematical model (fitted) in the microcontroller to derive several key parameters. These key parameters, along with user input information (weight, height, gastrointestinal-related questionnaire information, etc.), are processed by a trained machine learning model or other classification model, which is generated using KNN based on the (20 cases) patient data and built into the microcontroller to return classification results. Attached Figure Description

[0049] Figure 1 An embodiment of a breathalyzer device is shown. The front of the device is depicted with a screen and a mouthpiece support; the side of the device has an on / off button; and the mouthpiece.

[0050] Figure 2 A schematic diagram of the device in operation when used by a user is shown.

[0051] Figure 3 A schematic diagram illustrating the operation and wireless functions of the breathalyzer is shown.

[0052] Figure 4 An implementation scheme for the 3D-printed main body of a breathalyzer device is shown.

[0053] Figure 5 This diagram illustrates the operational phases of a breathalyzer device. Steps 1 through 7 are used to perform the breath test.

[0054] Figure 6 This is a schematic diagram of the lower half of a 3D-printed prototype of a dual-channel breathalyzer device. The diagram illustrates the channels and specifications of this embodiment. Other embodiments have different specifications and measurements.

[0055] Figure 7 A schematic diagram of the upper part of the breathalyzer is shown, while the lower part is depicted... Figure 6 middle.

[0056] Figure 8 The sensor bracket is shown, which is located at Figure 6 and Figure 7 At the end of each depicted channel. In some embodiments, the sensor holder contains a desiccant or filter; in other embodiments, the sensor holder does not contain a desiccant or filter.

[0057] Figure 9A schematic diagram of the upper part of a single-channel breathalyzer is shown, and a screen located on the upper part of the breathalyzer device is also depicted.

[0058] Figure 10 A schematic diagram of the lower half of a single-channel breathalyzer device is shown.

[0059] Figure 11 A schematic diagram of the assembled breathalyzer device, showing its upper and lower parts, is shown. In some embodiments, the 3D-printed breathalyzer device has two parts, an upper and a lower section, which complement each other and are sealed during assembly. The figure also shows the battery cover located on the lower section.

[0060] Figure 12 The chemical structure of polyaniline is shown.

[0061] Figure 13 As an example of a sensor, in this embodiment, it is a PANI-CSA sensor. Contact pads electrically connected to the circuitry are depicted, along with interdigitated fingers serving as electrodes, which can be gold or platinum.

[0062] Figure 14 The sensitivity of the PANI-CSA sensor to various gases is shown. This figure demonstrates the sensor's high sensitivity to ammonia.

[0063] Figure 15 The sensor mounting base and its specifications are shown. Figure 14 In one embodiment, the sensor is mounted using interdigitated electrodes. In other embodiments, the PANI-CSA can be placed on a mounting base without electrodes.

[0064] Figure 16 The resistivity of the PANI-CSA sensor for different concentrations of ammonia is shown. As illustrated, the resistivity changes even at ammonia concentrations below 25 ppb.

[0065] Figure 17 The diagram shows how current changes with resistivity. The higher the resistivity, the lower the current.

[0066] Figure 18 The resistivity change of the PANI-CSA sensor is shown in the presence of a sieve (a desiccant). The presence of the sieve reduces the resistivity by 50% due to the absorption of ammonia. This figure demonstrates that, in this embodiment, the ammonia sensor exhibits higher sensitivity as a measure of resistivity when the sieve is absent.

[0067] Figure 19The relationship between signals from an ammonia sensor and meals is illustrated. The graph depicts the ammonia content in the exhalation of users who are fasting (abstaining from food or drink except water for 8 to 12 hours), users between meals, and users 30 minutes after a meal. According to the graph and this embodiment, the ammonia resistivity in the exhalation is highest after 8 to 12 hours of fasting.

[0068] Figure 20 The calibration of the SPANI sensor is shown. The SPANI sensor is sensitive to CO2 concentrations from 0.5% to 3%.

[0069] Figure 21 An embodiment of a multianalyte breath tester is shown, which has an attached but removable mouthpiece, a touchscreen display covering the entire front of the device, and a power button.

[0070] Figure 22 An embodiment of the internal structure of a multianalyte breathalyzer device is shown, featuring 21 sensors. The device can accommodate more sensors as needed. This version of the device has an opening that serves as a mouthpiece and receives a breath sample from a user who surrounds their mouth with the opening. The opening accommodates several desiccant crystals embedded in its walls, but the mouth does not come into contact with these crystals. The device internally houses electronic circuitry and a microprocessor connected to each sensor, along with Bluetooth, a USB port, a battery, and a small orifice for expelling the breath as it leaves the device.

[0071] Figure 23 The exterior of the multi-sensor, multi-analyte device is shown.

[0072] Figure 24 The interior of a version of a multianalyte breath analyzer device is shown, which houses seven sensors. For 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) that indicate the presence of inflammatory bowel disease (IBD).

[0073] Figure 25 A schematic diagram of the interior of the dual-sensor device is shown.

[0074] Figure 26 A schematic diagram of the interior of the seven-sensor device is shown.

[0075] Figure 27 The screen of the multi-sensor, multi-analyte breath test device is shown, which has a drop-down menu and various diseases that the breath test device of this embodiment can detect.

[0076] Figure 28The results show that PANI / DNNSA has high sensitivity to two concentrations of hydrogen (20 kppm and 6 kppm).

[0077] Figure 29 Principal component analysis (PCA) of expiratory hydrogen detection data from individuals suspected of having celiac disease is presented. This PCA method reveals clusters of individuals testing positive for celiac disease in their expiratory breath tests.

[0078] Figure 30 This demonstrates SPANI's high sensitivity to CO2.

[0079] Figure 31 This demonstrates the ability of a breathalyzer device to use artificial intelligence—machine learning—to analyze breath test data. The machine learning algorithm, embedded within the breathalyzer device's Arduino microprocessor, yields results with accuracy superior to PCA or standard statistical analysis.

[0080] Figure 32 This demonstrates the ability of machine learning algorithms to better analyze data obtained through breath testing using a breathalyzer device.

[0081] Figure 33 The results showed that the accuracy rate of correct diagnosis reached 100% among users who were considered to have symptoms consistent with a certain disease when using the breathalyzer device.

[0082] Figure 34 The locations of the intervention (steps 6 and 7) are shown, along with the insertion of a learned file containing the algorithm into the microprocessor for analysis of user data from the breathalyzer device of this embodiment. In step 6, the retrieved data is processed by a mathematical model encoded in the microcontroller (fitted) to generate several key parameters. These key parameters, along with user input information (weight, height, gastrointestinal-related questionnaires, etc.), are processed by a trained machine learning model or other classification model based on data from (20 cases) of patients. KNN It generates and integrates into the microcontroller, and returns the classification results.

[0083] Figure 35 A schematic diagram of a ring-shaped tubular channel is shown, with multiple sensors positioned around its perimeter. Exhaled air enters through a sealed opening in the channel, at which a mouthpiece is securely attached. Detailed Implementation

[0084] The following detailed description should be read in conjunction with the accompanying drawings, in which the same elements in different drawings have the same reference numerals. The drawings are not necessarily drawn to scale, depict selected embodiments, and are not intended to limit the scope of the invention. Those skilled in the art will recognize that the examples provided herein have many available alternatives that fall within the scope of the invention.

[0085] In some embodiments, the present invention provides an improved breath test device and method for determining the presence of Helicobacter pylori in the digestive tract of a subject. This improved breath test device and method are less expensive, easier to use, and more accurate in diagnosis than existing methods and apparatus.

[0086] Exhaled human breath can contain up to 100 times more CO2 than inhaled air. This exhaled CO2 comes from multiple sources within the body, one of which may be Helicobacter pylori present in the gastrointestinal tract. In healthy individuals not infected with Helicobacter pylori, exhaled human breath contains approximately 3.8% carbon dioxide. 13 C is a naturally occurring isotope of carbon. 12 C is a more stable carbon-12 isotope and is found in CO2. 12 C is abundant in nature, making up 98.9% of the total carbon content; it exists in... 13 CO2 13 C is relatively rare in nature and accounts for only 1.1% of natural carbon.

[0087] Helicobacter pylori is a genotype-diverse bacterium that can alter its genetic makeup and mutate in vivo during colonization of the digestive tract of human subjects. The risk of developing gastric cancer is increased when Helicobacter pylori is positive for the cytotoxin-associated gene A (CagA) compared to when it is negative for CagA. In the Western world, the seropositivity rate for CagA in Helicobacter pylori is approximately 60%, compared to nearly 100% in Asian countries and much of Africa. The highly immunogenic CagA protein encoded by the CagA gene triggers a serum antibody response that can be detected by enzyme-linked immunosorbent assay (ELISA).

[0088] Despite their different genetic makeup, all types of Helicobacter pylori can use their abundant urease to process urea (whether or not labeled as...) 13 C) Hydrolysis to produce CO2 (or 13 CO2 and ammonia (NH3) gas. Once produced, ammonia and carbon dioxide (CO2 or...) 13 CO2 diffuses into the bloodstream through the gastrointestinal mucosa and is expelled from the lungs with exhaled air. The hydrolysis of urea, in both labeled and unlabeled forms, is illustrated by the following equations 1 and 2:

[0089]

[0090]

[0091] Previous urea breath tests required a labeled urea substrate to detect the amount of labeled CO2 in the breath. In contrast, the breath tester and method of the present invention use one or more sensors to detect the total amount of both ammonia and carbon dioxide present in the breath sample. Advantageously, the breath tester and method of the present invention do not require a labeled substrate.

[0092] Refer to the accompanying drawings, and in particular Figure 1 , Figure 2 and Figure 9 The figures show schematic diagrams of a breathalyzer according to the present disclosure. In some embodiments, the breathalyzer includes some or all of the components depicted in these figures; in other embodiments, the breathalyzer includes additional components beyond those depicted in these figures.

[0093] Some embodiments of a breathalyzer detect the presence and concentration of both ammonia and carbon dioxide in a breath sample. As described in more detail below, the breathalyzer includes an input, one or more gas sensors, circuitry, and a processor. The input receives a breath sample. At least one gas sensor is in contact with the breath sample. At least one of the one or more gas sensors comprises an ammonia-selective material (e.g., doped polyaniline) and a conductive material. The circuitry operatively connects the conductive material to the processor. The processor detects changes in resistivity in the circuitry and uses these changes in resistivity to calculate the concentration of ammonia in the breath sample. The same or different gas sensors comprise a carbon dioxide-selective material (e.g., doped polypyrrole) and a conductive material. The circuitry operatively connects this conductive material to the processor. The processor detects changes in resistivity in the circuitry and uses these changes in resistivity to calculate the total concentration of carbon dioxide in the breath sample.

[0094] The breathalyzer includes a mouthpiece with an open end to allow a breath sample to move through it. The mouthpiece can be made of any suitable type of material, including but not limited to plastic or metal. The mouthpiece includes a first part and a second part. The first part is configured as an input end to receive a breath sample. The size and shape of the first part can be designed to accommodate the user's lips, allowing the user to blow exhaled breath into the mouthpiece.

[0095] In some cases, the mouthpiece includes a one-way valve. In such cases, the user exhales into the mouthpiece. The exhaled air moves forward through the one-way valve and is captured. In other words, the exhaled air cannot move backward through the one-way valve and enter the first part.

[0096] The breathalyzer also includes a main body. A mouthpiece is attached to the main body. Specifically, a second portion of the mouthpiece is sized and shaped to be removably attached to the main body. For example, this second portion can be snap-on or possibly threaded onto the main body. The main body can contain plastic, metal, or any other suitable material. In some cases, the main body and mouthpiece contain the same material; in others, they contain different materials.

[0097] In some implementations, the mouthpiece is a disposable mouthpiece. Disposable mouthpieces are ideal because they can be replaced for each new user. Furthermore, in some cases, components of the mouthpiece and / or body that come into contact with exhaled breath may be made of an inert or non-reactive material (e.g., polytetrafluoroethylene (PTFE)) that does not interfere with ammonia absorption.

[0098] The mouthpiece can be integrally formed with the body, or it can be a separate structure connected to the body. In the example where the mouthpiece is a separate structure connected to the body, the mouthpiece can be placed inside the body through a hole in the body, for example, by pushing, screwing, or pressing.

[0099] In one implementation, the mouthpiece is permanently attached to the body. In such instances, the mouthpiece may be securely mounted to the body, extending straight out from the body or at an angle. These alternative configurations allow the eyes of the subject taking the breath sample to be either directly facing or away from the body during the sampling process. The mouthpiece may be permanently attached to an opening in the body using a support made of plastic, metal, or any other material. In other cases, the mouthpiece may be permanently attached to the body without the use of a support.

[0100] Figure 10 and Figure 11 An exemplary implementation of a breathalyzer is illustrated. Figure 10 A breathalyzer with a mouthpiece attached to the main body is shown, while Figure 11 A breathalyzer with a mouthpiece detachable from the main body is shown. In these embodiments, the breathalyzer is provided as a stand-alone, portable handheld device. In some cases, the mouthpiece is a disposable mouthpiece that is discarded after use and can be replaced with a new mouthpiece for each new user.

[0101] The mouthpiece can be attached to the exterior of the breathalyzer body or can extend into the breathalyzer body. The mouthpiece can be attached to any location on or within the breathalyzer, provided that the first end of the mouthpiece can reach the lips of the subject taking the breath test. The mouthpiece can be attached to the breathalyzer via any suitable type of connection, including direct connection, push-in connection, or threaded connection, or another type of connection within the breathalyzer body. In some cases, the mouthpiece can be glued or can be adhered to the body using any other type of adhesive.

[0102] The mouthpiece can have any desired shape; for example, it can be oval, cylindrical, conical, or straw-shaped. The shape of the mouthpiece should allow the subject's lips to fit snugly around it. As disclosed above, the mouthpiece may optionally include a self-sealing one-way valve to isolate the exhaled sample from the surrounding air once it leaves the mouthpiece and enters the body of the breath analyzer.

[0103] The mouthpiece may optionally include a lining material positioned inside the mouthpiece. When installed, this lining material covers part or all of the inner surface of the mouthpiece. In some cases, the lining material is a desiccant capable of trapping moisture. The desiccant may contain silica, activated carbon, calcium sulfate, calcium chloride, or any other type of desiccant (composed of or substantially composed of these). The desiccant may also contain any or a combination of these or other desiccants. The desiccant may optionally have a color indicator to indicate the amount of moisture trapped by the desiccant. In other instances, the mouthpiece does not contain any type of desiccant or other lining material.

[0104] In another embodiment, the mouthpiece includes a liner material configured to trap and absorb some of the gases contained in the exhaled breath sample while allowing other gases to pass through. For example, the liner material on the mouthpiece can block H2O, nitric oxide (NO), methane (CH4), nitrogen (N2), and other volatile organic compounds contained in human exhalation, while allowing CO2 (or...) 13 CO2 and NH3 pass through the lining material and through the nozzle.

[0105] Breathalyzers may optionally include one or more filters (or “traps”). These filters perform a similar function to the mouthpiece liner material; that is, they allow certain gases to pass through while trapping and preventing others from passing. Materials used for the filters may include sodium hydroxide, silica, activated carbon, calcium sulfate, calcium chloride, or any other type of desiccant. Filters may also include humidity sensors, such as hygrometers. However, in some cases, breathalyzers do not include any filters.

[0106] In one embodiment, the filter is a single filter configured to block the passage of specific gases present in human exhalation that are not intended to be measured by a breathalyzer. For example, the filter may block moisture, nitric oxide, methane, oxygen, and / or other volatile organic compounds in the exhalation sample. This single filter allows ammonia (NH3), CO2, and... 13 The passage of CO2, or only ammonia, only CO2, or only [other substances] are permitted. 13 CO2 passes through.

[0107] In another embodiment, the filter comprises multiple filters. These filters can have any desired shape and can contain various types of materials. The shape of the filters is not limited and can be circular, oval, square, or a combination of different shapes. Each of these filters can trap one or more undesired gases (i.e., those gases not intended to be measured) and allow CO2, 13 CO2 and ammonia pass through, either alone or together.

[0108] The breathalyzer includes at least one gas sensor, a processor, and a power supply. The at least one sensor, processor, and power supply are electrically connected via circuitry.

[0109] The processor can be any processor known in the art. In some cases, the processor is a microcontroller. In other cases, the processor is an Arduino microcontroller.

[0110] At least one sensor may have any of the embodiments described. In some cases, the at least one sensor includes... Figure 13 The sensor is shown in the embodiment. The at least one gas sensor is electrically connected to the circuit using any desired connection mechanism. In some cases, the at least one sensor is connected to the circuit via an optional sensor mount. In such cases, the at least one sensor can be directly mounted to the sensor mount. The sensor mount serves as an interface between the sensor and the circuit. Therefore, the sensor mount can be any structure known in the art for connecting the electrodes of at least one sensor to the circuit. In some embodiments, the sensor mount is a printed circuit board.

[0111] In other cases, at least one sensor is directly connected to circuit 4. For example, in some embodiments, the circuit includes two metal clips that can be clamped onto contact pads to form an electrical connection. A user can also replace an old sensor with a new one by removing the old sensor from the metal clips and inserting the new sensor into the clips.

[0112] The main body also includes an on / off button and a power supply. The power supply can be a portable power source, such as a battery. When the on / off button is activated, the power is switched on. For example, as shown, the power supply provides voltage to a regulator. In some cases, the power supply provides a voltage of a certain number of volts to the regulator. The regulator adjusts the magnitude of the voltage sent to the sensor. In some cases, the regulator provides a voltage of 0 to 5 volts to at least one sensor. In some cases, the regulator provides a voltage of approximately 5 volts to at least one sensor. In one embodiment, the regulator is an IC1 7805 regulator manufactured by Fairchild Electronics.

[0113] A resistor is also electrically connected to at least one sensor and provides resistance to it. In some cases, this resistor is 10 kΩ. When a breath sample comes into contact with at least one sensor, the resistivity of the sensor changes, and this change is related to the amount of ammonia and / or carbon dioxide present in the sample. At least one sensor outputs a voltage (and the change in resistivity) to a processor. The processor detects the change in resistivity in the at least one sensor and uses these changes to calculate the concentrations of ammonia and carbon dioxide in the breath sample. The processor can also compare the ammonia concentration between two different breath samples. Similarly, the processor can also compare the carbon dioxide concentration between two different breath samples.

[0114] Gas sensors may include a single gas sensor or multiple gas sensors. Each gas sensor is configured to detect and measure ammonia, CO2, and [other gases] in a human exhaled breath sample. 13 One or more of CO2. Therefore, at least one gas sensor 50 is sensitive to ammonia (NH3), at least one gas sensor is sensitive to CO2, and at least one gas sensor is sensitive to... 13 CO2 sensitive. For example, one gas sensor can be sensitive to ammonia, another gas sensor can be sensitive to CO2, and yet another different gas sensor can be sensitive to... 13 Sensitive to CO2. Alternatively, a single gas sensor can be sensitive to any combination of these gases. For example, a gas sensor can be sensitive to ammonia as well as CO2 and... 13 CO2 sensitive; another gas sensor can be sensitive to ammonia (and also to CO2 or CO2). 13 (not sensitive to CO2); and another gas sensor can detect both CO2 and... 13 CO2 sensitive (and insensitive to ammonia). One or more gas sensors can measure humidity in exhaled breath to assess the effect of humidity on the sensitivity of the gas sensor to a specific gas during detection.

[0115] Breathalyzers can measure ammonia and CO2 simultaneously or sequentially (including...) 12 CO2,13 CO2, and / or 12 CO2 and 13 (Both CO2 and CO2). When the breathalyzer 10 measures ammonia and CO2 sequentially, the two measurements occur within a short time interval (e.g., about 1 to 10 seconds) between each other. This allows the breathalyzer to measure and process the effects of each gas sensor simultaneously or nearly (i.e., substantially) simultaneously.

[0116] Each gas sensor comprises a substrate, a conductive material, and a gas-selective material. The conductive material is deposited onto the substrate and can contain or consist of any desired conductive material. In some cases, the conductive material is platinum; in others, it is gold.

[0117] In some embodiments, the conductive material is an electrode arrangement. This electrode arrangement can be a single electrode or multiple electrodes. The electrodes can be spaced apart in any desired arrangement. In some cases, the electrode spacing is less than about 250 μm, possibly less than about 150 μm, such as about 100 μm. In some cases, the electrode spacing is less than about 10 μm, such as 5 μm. In some embodiments, such as... Figure 13 The gas sensor shown includes interdigitated electrodes. In one embodiment, the gas sensor includes platinum interdigitated electrodes with a line spacing of 100 μm or less.

[0118] One or more gas sensors can operate at room temperature (i.e., 68℉ to 77℉, or 20℃ to 25℃) or at higher temperatures. One or more gas sensors can be manufactured by any suitable method. Such methods can include spin coating, drop coating, sol-gel methods, or any other known manufacturing method. One or more gas sensors can be manufactured for single use or multiple reuses. One or more gas sensors can contain polyaniline or polypyrrole doped with a protic acid. As a non-limiting example, one or more gas sensors can contain PANI / CSA (polyaniline / camphor sulfonic acid), PANI / DNNSA (polyaniline / dinonylnaphthalene sulfonic acid), or PPY / DBSA (polypyrrole / dodecylbenzene sulfonic acid). The applicant has found that the PANI / CSA gas sensor is sensitive to the detection of ammonia, and additionally, PANI / DNNSA and PPY / DBSA are sensitive to the detection of... 12 CO2 and / or 13 They are particularly sensitive to CO2.

[0119] One or more gas sensors may include those capable of simultaneously or sequentially sensing ammonia (NH3) and / or carbon dioxide (CO2 and / or other gases). 13One or more gas sensors may be thin-film or thick-film sensors for CO2. Such gas sensors may comprise polymer thin films, polymer thick films, or polymer composite films. One or more gas sensors may be metal oxide sensors, such as zinc oxide (ZnO) or other metal oxide sensors. One or more gas sensors may be blends of polymers with metal oxides (such as polyaniline, polypyrrole, or another polymer combined with zinc oxide or another metal oxide). In some cases, one or more gas sensors comprise blends of sulfonated polyaniline or polyethyleneimine (PEI) with a polyelectrolyte. In other cases, one or more gas sensors may be blends of emerald green imine-based polyaniline (EB-PANI) with poly(3,4-ethylenedioxythiophene)-polystyrene sulfonic acid (PEDOT:PSS).

[0120] In some cases, ammonia-selective materials contain doped polyaniline. Polyaniline exhibits three distinct oxidation states: colorless imine (LEB, fully reduced state), emerald green imine (EB, semi-oxidized state), and aniline black (PNB, fully oxidized state). Furthermore, when in the emerald green imine state, polyaniline can exist as either an emerald green imine salt or a emerald green imine base. In the emerald green imine salt form, polyaniline is conductive. The emerald green imine salt form is typically obtained by protonating a basic amine and imine site with a strong acid. This process is reversible because the emerald green imine base form is obtained by deprotonating the amine group. Therefore, the emerald green imine state of polyaniline transitions between acidic and basic forms.

[0121] The inventors discovered that the acid-base transition of polyaniline gives it pH sensitivity, and this property enables its effective use in ammonia detection. When ammonia comes into contact with the emerald green imine salt form of polyaniline, the ammonia causes the amine groups to undergo a deprotonation reaction, converting them into emerald green imine bases. This process also leads to an increase in resistivity and a corresponding decrease in conductivity.

[0122] In some cases, one or more gas sensors may contain dopants to enhance the sensitivity of gas sensor 50. For example, when at least one gas sensor 50 uses polyaniline, its pH sensitivity can be enhanced by doping the polyaniline with a protic acid. For ammonia detection, polyaniline with increased pH sensitivity is ideal because when ammonia converts polyaniline to a chlorophylline base, it results in an even greater increase in resistivity and a corresponding decrease in conductivity. The greater increase in resistivity (and the decrease in conductivity) is desirable because it is easier to detect, and thus improves the sensitivity of polyaniline to ammonia.

[0123] Polyaniline doped with protic acids exhibits improved pH sensitivity compared to undoped polyaniline. In some cases, polyaniline can be doped with ions such as Cl⁻ and SO₄²⁻. 2-The protic acid is used to obtain a pH sensitivity of approximately 59 mV. Certain protic acids result in even greater increases in pH sensitivity. For example, polyaniline doped with camphorsulfonic acid has shown a pH sensitivity of approximately 70 mV.

[0124] In some cases, polyaniline contains at least one dopant that enhances its pH sensitivity. In some cases, the dopant is a protic acid. In some embodiments, the dopant is hydrochloric acid. In other embodiments, the dopant is camphorsulfonic acid. In still other embodiments, the dopant is both hydrochloric acid and camphorsulfonic 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. Furthermore, in some cases, polyaniline has a dopant that provides a pH sensitivity greater than 59 mV. In one embodiment, polyaniline has a camphorsulfonic acid dopant that provides a pH sensitivity of approximately 70 mV, a higher sensitivity observed when using other dopants.

[0125] Furthermore, the gas-selective material of the gas sensor can be directly deposited onto the conductive material using any desired deposition process. For example, the gas-selective material can be deposited onto the conductive material using spin coating, drop coating, chemical vapor deposition, or sputtering. In some cases, the conductive material is coated with a spin-cast gas-selective material. In some embodiments, the gas-selective material is doped polyaniline deposited directly onto the conductive material using spin coating.

[0126] As discussed above, one or more gas sensors can detect and measure one, two, or more gases. At least one of the gas sensors detects ammonia (NH3) at very low ppb levels. In some cases, one or more gas sensors 50 detect ammonia concentration levels in the range of 1 ppb to 1 ppm. In some cases, one or more gas sensors 50 detect ammonia in concentration ranges of 10 ppb and above (e.g., in the range of about 10 ppb to about 50 ppb), or in concentration ranges of 500 ppb and above. In still other cases, one or more gas sensors 50 detect exhaled ammonia levels below 50 ppb and up to 500 ppm.

[0127] As noted above, at least one of one or more gas sensors detects CO2. The detected CO2 can be... 12 CO2, 13 CO2, or 13 CO2: 12The ratio of CO2. In some cases, at least one of the gas sensors 50 detects CO2 concentrations in the range of 10 ppm to 1500 ppm and above (or... 13 CO2) Figure 6 In some cases, at least one gas sensor detects CO2 levels as low as 1 mM / min or 50 ppm to 100 ppm. Furthermore, in some cases, at least one gas sensor detects CO2 levels as low as 1 mM / min or 50 ppm to 100 ppm. 13 CO2.

[0128] One or more gas sensors may comprise a thin-film polyaniline (PANI) doped with hydrochloric acid (HCl). This sensor can be prepared by chemically oxidizing and polymerizing aniline in an acidic aqueous medium (1 M HCl) using ammonium persulfate (APS) as the oxidant. PANI sensors may be doped with acid dopants other than HCl. In some cases, one or more gas sensors may comprise a thin-film polyaniline doped with camphorsulfonic acid (PANI-CSA) and also doped with HCl or another acid dopant.

[0129] In some cases, one or more gas sensors may comprise a thick-film polymer, pyrrole. Such sensors can be polymerized in the presence of an oxidizing agent (such as FeCl3 or any other acid). The polymerized pyrrole (polypyrrole) is capable of detecting and measuring CO2 concentrations of 10 ppm and above. 13 CO2.

[0130] In some cases, one or more gas sensors may include those capable of sensing ammonia, CO2, and / or... 13 A fiber optic sensor for CO2. Other gas sensors 50 besides those explicitly mentioned herein are envisioned as one or more gas sensors 50 for detecting and measuring ammonia, CO2, and... 13 CO2.

[0131] A non-limiting method for manufacturing a PANI-CSA thin-film sensor is as follows: The film can be prepared by spin-coating a PANI-CSA solution onto an interdigitated array (IDA) electrode. Prior to spin-coating, the electrode can be cleaned by rinsing in methanol, followed by rinsing with deionized water, and drying in a dry nitrogen stream. The PANI film can then be spin-cast onto the IDA electrode by adding 100 μl of solution at 500 RPM. The electrode pads can be cleaned, for example, with a cotton swab dipped in methanol to facilitate direct electrical contact with the breathalyzer.

[0132] The breathalyzer also includes contact pads and electronic circuitry 60. One or more gas sensors are connected to the electronic circuitry via the contact pads. The electronic circuitry is electrically connected to the one or more gas sensors. The electronic circuitry can be as follows: Figure 5 The work shown, or can be done as follows Figure 23 The circuitry is capable of recording multiple consecutive values ​​for each measured gas detected by one or more gas sensors. In some cases, the circuitry records the highest detected value or average value for each gas detected by one or more gas sensors. Using software, the circuitry converts the measured resistivity of the gas into concentration (ammonia in ppb, and...). 13 CO2 and CO2 are expressed in ppm.

[0133] In some embodiments, the breathalyzer includes a display. The display is electrically connected to a processor. The display is configured to visually display ammonia, CO2, and [other gases detected by one or more gas sensors]. 13 The amount of CO2 gas is measured. The display can be a window display mounted on the main body of the breathalyzer. In an alternative embodiment, the results can be displayed on a computer portal or other fixed or portable device using Bluetooth technology or any other wireless data transmission device.

[0134] The display can show representations of all three gases (ammonia, ...). 13 A single result of the summation of CO2 and CO2, or a result that can be shown for each corresponding gas (i.e., one output for ammonia, one output for CO2, and one output for CO2). 13 CO2 is one output. In some implementations, the results are displayed numerically (e.g., from 1 to 1000, where a value of 1 indicates no Helicobacter pylori infection and a value of 1000 indicates that the subject was infected with Helicobacter pylori), and / or in the actual units of the measured gas (e.g., ammonia concentration in ppb, CO2 and / or 13 (CO2 concentration in ppm). In other embodiments, the results may be displayed as a "yes" or "no" indication, and / or presented using a light source with color. For example, a green light on display 30 may indicate a negative test result, and a red light on display may indicate a positive test result. Any or more of these results (e.g., numerical value, actual unit of the gas measured, color, yes / no indication) may be displayed on the screen.

[0135] In some embodiments, the body is configured to include a first compartment and a second compartment. In some cases, the first compartment is a chamber housing at least one gas sensor and a sensor mounting base, and the second compartment is an electrical enclosure housing various components. In some embodiments (not shown), each of the at least one gas sensor may be provided with a separate chamber.

[0136] In some cases, the chamber includes an openable and closable lid or door. When the door is closed, the chamber provides a closed, sealed environment around at least one sensor; when the door is open, at least one gas sensor is accessible through the door opening. The user can open the chamber door as needed to remove and replace at least one sensor. The chamber also includes an outlet. The outlet includes a lid that can be opened to release an exhaled breath sample from the chamber, or it can be closed to trap the exhaled breath sample within the chamber.

[0137] The mouthpiece connects to the chamber, allowing exhaled air to flow directly into the chamber from the mouthpiece. As discussed above, the breath detector 10 may include a filter. In some cases, this filter is a desiccant assembly. The desiccant assembly helps remove excess moisture from the exhaled air. In some embodiments, the desiccant assembly is disposed inside the mouthpiece. In other embodiments, the desiccant assembly is disposed inside the chamber. The exhaled air first moves through the desiccant assembly and then comes into contact with at least one gas sensor.

[0138] In some cases, the desiccant assembly is provided in the form of a tube through which exhaled air flows. The tube may have an interior filled with a plurality of desiccant beads. These desiccants may also be arranged to form multiple channels for exhaled air to flow through. In this embodiment, the tube may have an interior filled with a plurality of desiccant beads 64 arranged to form a single channel.

[0139] As discussed above, the breathalyzer may include one or more optional gas filters 13. The gas filters help remove selected gases (e.g., nitrogen and / or hydrogen) from the exhaled breath. In some embodiments, the gas filter is located inside the mouthpiece. In other embodiments, the gas filter is located inside the chamber. The exhaled breath first moves through the gas filter and then comes into contact with the sensor.

[0140] The main body may also include a second compartment configured to house an electrical enclosure for various components. In the illustrated embodiment, the electrical enclosure houses a processor, some circuitry, a power supply, a display, and a wireless connector.

[0141] During use, the user activates the breathalyzer by turning on / off the switch. This switch can be located anywhere near the external surface of the main body. The switch 26 then causes the power supply to provide voltage to the voltage regulator. The voltage regulator regulates and provides voltage to at least one gas sensor. A resistor also provides resistance to at least one gas sensor.

[0142] The user then blows a breath sample into the first part of the mouthpiece. The breath sample moves through a one-way valve and across an optional desiccant / gas filter. The breath sample then exits the optional desiccant / gas filter and enters a chamber where it comes into contact with at least one gas sensor. The breath sample causes a change in resistivity in the at least one gas sensor, and this change in resistivity is output to a processor.

[0143] This disclosure also provides a urea breath test (“UBT”) method, which is an improvement on existing urea breath test methods. This novel UBT uses the breath analyzer described above, which is simple to operate, non-invasive, inexpensive, and substantially risk-free. The breath analyzer and related breath test method can be used by patients at the point of care for self-testing, and the test results are available immediately.

[0144] To conduct this novel UBT, all subjects wishing to be tested for Helicobacter pylori infection must avoid ingesting antibiotics, bismuth preparations, sucralfate, and proton pump inhibitors for two weeks prior to using the breath tester and the breath test method.

[0145] On the day of the breath test, the subject will follow the procedure below:

[0146] In one implementation, the subject fasts and abstains from drinking for one hour before exhaling into the breathalyzer. At the end of the one-hour fast, the subject places their lips to the mouthpiece and exhales for the first time, completing a sustained exhalation lasting approximately 5 seconds or between 3 and 8 seconds, depending on the subject's age, height, and weight, delivering the exhaled air through the mouthpiece into the breathalyzer. This provides a baseline exhalation.

[0147] Following the first exhalation, the subject will ingest urea in an amount approximately 5 mg / kg of body weight, or a standard dose of 300 mg, or another standard dose of 125 mg or 250 mg, or another single or multiple dose depending on the subject's age, weight, and general clinical condition. The ingested urea can be in two forms. It can be labeled urea (…). 13 Urea can be taken in either CO(NH2)2 or unlabeled urea (CO(NH2)2). Any dose of urea can be taken in capsule form with a small amount of water, or mixed into small portions of food, such as solid food (e.g., pudding) or liquid (e.g., lemon juice) and taken orally.

[0148] Within a span of 10 to 30 minutes after ingesting urea (capsule or urea-containing meal), the subject will exhale a second breath into the device via a mouthpiece. This exhalation via the mouthpiece can be performed at any time between 10 and 30 minutes after urea ingestion. For example, the exhalation could be 10 minutes, 15 minutes, 20 minutes, or 30 minutes after urea ingestion (whether urea is labeled or unlabeled). In this embodiment, the second exhalation must not be earlier than 10 minutes after urea ingestion (whether in capsule form or urea-containing meal) and must not be later than 100 minutes after urea ingestion (whether in capsule form or urea-containing meal). Typically, the second exhalation occurs 15 to 30 minutes after ingestion of urea or a urea-containing meal.

[0149] In an alternative implementation, the subject ingests a high-protein meal (e.g., a hamburger, high-protein bar, or equivalent) after completing a baseline exhalation. The subject then exhales a second time into mouthpiece 12 within a span of 20 to 120 minutes after ingesting the high-protein meal. This second exhalation can occur at 20 minutes, 30 minutes, 50 minutes, 80 minutes, and 120 minutes after ingesting the high-protein meal. Typically, the second exhalation occurs 60 minutes after ingesting the high-protein meal.

[0150] Exhaled breath (baseline and post-urea intake) enters the main body of the breathalyzer through a filter (if any), or directly through the mouthpiece if no filter is present. Each exhaled breath exits the filter (or mouthpiece) and enters one or more gas sensors. As the exhaled breath sample (baseline exhalation and post-urea intake exhalation) enters one or more gas sensors in the breathalyzer, the resistivity of one or more gas sensors will vary with the amount of ammonia and / or CO2 and / or CO2 in the detected exhaled breath. 13 The amount of CO2 changes.

[0151] When at least one gas sensor is a single gas sensor, the sensor can detect three gases: ammonia, CO2, and... 13 CO2. When at least one gas sensor includes two gas sensors, one of which can detect ammonia, and the other gas sensor can detect CO2 and / or 13 CO2. When at least one gas sensor includes three sensors, one gas sensor can detect ammonia, one gas sensor can detect CO2, and another gas sensor can detect... 13 CO2. When at least one gas sensor includes four gas sensors, one gas sensor can detect ammonia, one different gas sensor can detect CO2, and another gas sensor can detect... 13CO2. When at least one gas sensor includes four gas sensors, one gas sensor can detect ammonia, another gas sensor can detect CO2, and yet another gas sensor can detect... 13 It detects CO2 and also has a sensor to detect moisture.

[0152] The resistivity change caused by the presence of ammonia in a breath sample was converted into an electric current, and then into parts per billion (ppb) of ammonia. Similarly, CO2 or 13 The resistivity change caused by the presence of CO2 is converted into current, and then into parts per million (ppm). The resistivity change (ΔR) (value after urea intake minus baseline value) is converted into ppb and ppm, and any measurement of the gas combination is performed using software specifically developed for the functions and electronics of the breathalyzer.

[0153] In one implementation, the display will show the ammonia concentration in ppb and the CO2 (and / or) concentration. 13 CO2 and / or 13 CO2 (CO2) values ​​expressed in ppm. In this embodiment, these values ​​corresponding to gas concentrations will be displayed through a window on the main body of the breathalyzer.

[0154] In another embodiment, the value on display 30 will be a single value, calculated by relating the amount of ammonia in the subject's baseline exhalation and exhalation after urea ingestion to the amount of CO2 or... 13 CO2 or 13 CO2: The amount of CO2. This calculation can be performed based on a linear prediction equation for the outcome. For example, the equation can have the following form:

[0155] (Ammonia logarithm to base 10 × ammonia constant) + (CO2 × CO2 constant) or

[0156] (The logarithm of ammonia to base 10 × the constant of ammonia) + ( 13 CO2× 13 (CO2 constant) or

[0157] (Ammonia logarithm to base 10 × ammonia constant) + (CO2 × CO2 constant) + ( 13 CO2 × 13 (CO2 constant) or

[0158] (The logarithm of ammonia to base 10 × the constant of ammonia) + ( 13 CO2:CO2 × 13 CO2: the constant of CO2) or

[0159] Another polynomial equation form, where ammonia is expressed in ppb, and 13 CO2 and CO2 are expressed in ppm, and 13 CO2: CO2 is expressed numerically.

[0160] This constant is obtained by analyzing ammonia, CO2, and... in various populations at risk of Helicobacter pylori infection and the general population. 13 The CO2 values ​​were obtained from multiple sets of data, as well as from analyses of various dietary habits in this population and the general population (e.g., regression analysis). This equation will be embedded in the software.

[0161] The linear prediction equation for the outcome can have the following basic form:

[0162] f(i) = β0 + β 1xi1 + ... + β p x ip , where i is a data point.

[0163] When the value obtained from the equation is below a certain threshold, the subject is considered negative for Helicobacter pylori infection. When the value obtained from the equation is above a certain threshold, the subject is considered positive for active Helicobacter pylori infection. A few inconclusive results will require repeat testing, and if the result remains inconclusive, the subject will be further evaluated using additional methods.

[0164] As discussed above, CO2 (or...) is produced by the hydrolysis of urea (unlabeled or labeled) by urease produced by Helicobacter pylori. 13 CO2 and ammonia diffuse into the bloodstream via the mucous membranes and are expelled from the lungs through exhaled air. Infection subjects tested using this invention ingesting labeled or unlabeled urea will produce elevated levels of CO2 (or ammonia) in their exhaled breath. 13 CO2 and ammonia.

[0165] As discussed above, subjects using this method are required to exhale twice into the breathalyzer. The first (baseline) breath is taken one hour after fasting and abstaining from water, and the second (after urea ingestion) breath is taken after ingesting urea (labeled or unlabeled) or a high-protein meal or high-protein bar. Throughout the method, the breathalyzer 10 will detect and measure ammonia, CO2, and / or... 13 The amount of CO2.

[0166] When urea is ingested, the ammonia, CO2, and / or exhaled ammonia after urea is 13 When CO2 levels are higher than baseline levels, and the increase is greater than 100% of baseline levels, the tested subject can be diagnosed with Helicobacter pylori infection. Additionally, when urea is ingested, the exhaled ammonia and CO2 (and / or...) 13When the total CO2 level is higher than the baseline level, and the increase is greater than 50% of the baseline level, the subject can be diagnosed with Helicobacter pylori infection.

[0167] In diagnosing Helicobacter pylori infection, the method disclosed in this invention has higher sensitivity and specificity than previous methods. This is because the method disclosed in this invention detects ammonia and CO2 (and / or...) 13 The total amount of CO2, rather than independent of CO2 (and / or 13 CO2 detection of ammonia, or CO2 detection independently of ammonia (and / or) 13 CO2). Because this method measures all products generated from urea hydrolysis (including ammonia, CO2). 12 CO2 and 13 CO2), therefore, the method disclosed herein provides the most accurate method for detecting and diagnosing Helicobacter pylori infection. Simultaneously, it detects CO2 (or...) in exhaled breath. 13 C O2 Measuring CO2 (or CO2) significantly minimizes the impact of not detecting ammonia when measuring CO2 (or CO2) and ammonia. 13 CO2), or CO2 not detected (or 13 Statistical and other errors may occur when measuring ammonia (CO2).

[0168] ammonia and CO2 (and / or) levels in subjects infected with Helicobacter pylori 13 The combined increase in CO2 is multiplicative (or exponential), rather than additive. Therefore, even a small difference between baseline exhalation and exhalation after urea ingestion can diagnose Helicobacter pylori infection, thus significantly reducing false negative results.

[0169] In infected subjects, ammonia and CO2 (and / or 13 The combined increase of ammonia and CO2 is more specific and sensitive for Helicobacter pylori infection than existing breath test methods because ammonia and CO2 (and / or CO2) are more effective against it. 13 The combination of CO2 can only suggest and diagnose Helicobacter pylori infection in individuals without end-stage liver disease, encephalopathy, end-stage renal disease, and metabolic diseases (such as Krebs cycle defect, urea cycle defect) (but cannot diagnose other infections).

[0170] The following non-limiting embodiments are intended to demonstrate how the breath testing method and breath testing device of this disclosure can be used to detect Helicobacter pylori infection in humans, adults and children.

[0171] Example 1 - After ingesting unlabeled urea (CO(NH2)2), the increase in ammonia and carbon dioxide in exhaled breath was calculated to diagnose Helicobacter pylori infection.

[0172] Subjects avoided antibiotics, bismuth preparations, proton pump inhibitors, and sucralfate for two weeks. After the two-week period, subjects fasted for one hour. At the end of the one-hour fast, subjects exhaled air into a breathalyzer via a mouthpiece. The exhalation lasted approximately 2 to 10 seconds. In another implementation, the exhalation continued until the subject emitted a specific sound or light, indicating that a sufficient amount of exhaled air had entered the subject. "Sufficient" means that the exhaled air sample was sufficient to come into contact with one or more gas sensors.

[0173] As baseline exhaled breath enters the body, the exhaled breath sample passes through a filter (when the filter is installed in place). Gases such as NO, CH4, N2, volatile organic compounds, O2, and moisture are blocked from reaching at least one gas sensor. Ammonia and CO2 are not blocked by the filter and are allowed to come into contact with one or more gas sensors.

[0174] When a breath sample is introduced into one or more gas sensors, the resistivity of these sensors changes according to the amount of ammonia and CO2 in the exhaled breath. Using electronic circuitry and software, the resistivity of at least one gas sensor is converted into ppb of ammonia and ppm of CO2. The ammonia and CO2 values ​​obtained during the first exhalation can be stored in memory.

[0175] At the end of the baseline exhalation, the subject ingests a known amount (e.g., 125 mg, 250 mg, or 300 mg, depending on the subject's weight and age) of unlabeled urea, in the form of a tablet, small meal, or beverage. Within a span of 10 to 120 minutes after ingestion of the unlabeled urea, the subject exhales a second time through the mouthpiece following the same procedure as the first exhalation to perform a post-urea exhalation. Typically, the post-urea exhalation occurs 20 minutes after ingestion of the unlabeled urea. The post-urea exhalation does not occur earlier than 10 minutes or later than 120 minutes after ingestion of the unlabeled urea. The ammonia (ppb) and CO2 (ppm) values ​​in the second exhalation are also stored in the memory of the breathalyzer device 10.

[0176] The breathalyzer will calculate the final values ​​of ammonia and CO2 as follows:

[0177] Final ammonia = Ammonia after urea intake minus baseline ammonia

[0178] Final CO2 = CO2 after urea intake minus baseline CO2.

[0179] The software can be used to convert resistivity values ​​into final ammonia concentration (ppb) and final CO2 concentration (ppm). In subjects testing positive for Helicobacter pylori infection, baseline ammonia levels will range from 20 ppb to 200 ppb, and post-urea ingestion ammonia levels will range from 80 ppb to 600 ppb. Because there is overlap between baseline and post-urea ingestion ammonia levels, the actual percentage change between baseline and post-urea ingestion ammonia will be used as a marker for Helicobacter pylori infection. A percentage change greater than 200% between baseline and post-urea ingestion ammonia is considered positive for Helicobacter pylori infection. A percentage change less than 200% is considered negative for Helicobacter pylori infection.

[0180] In subjects who test positive for Helicobacter pylori, baseline CO2 concentrations are typically below 200 ppm, and CO2 concentrations after urea ingestion are typically above 200 ppm (e.g., ranging from 200 ppm to 1000 ppm). A subject is considered positive for Helicobacter pylori infection when the percentage change between the calculated baseline CO2 and the CO2 after urea ingestion is greater than 1000%.

[0181] In one implementation, a positive test result will be a percentage change in ammonia greater than 200% and a percentage change in CO2 greater than 1000%; expressed as numerical values, ppb, and ppm, respectively, as determined using device-specific software.

[0182] In another implementation, the device software converts the final result into a numerical value recognizable by a computer program. This final result is calculated using an equation that considers the percentage or actual increase in ammonia and the percentage or actual increase in CO2 between a baseline breath sample and a breath sample after urea ingestion. This equation can be logarithmic or statistical and includes values ​​for two variables (ammonia and CO2) corresponding to the tested subject, two constants, and one or more coefficients. These constants and coefficients are calculated based on the epidemiological and individual characteristics of the subject infected with Helicobacter pylori, such as age, sex, race, height, and weight.

[0183] In another embodiment, a positive result for Helicobacter pylori will be represented by one color (e.g., red), and a negative result for Helicobacter pylori will be represented by one color (e.g., green). In another embodiment, a positive result for Helicobacter pylori infection will be represented by a plus sign (+), and a negative result for Helicobacter pylori infection will be represented by a minus sign (−).

[0184] The final results can be displayed through the window on the main body of the breathalyzer. The final results (whether positive or negative, yes / no, different colored indicator lights, or ammonia and CO2 values) will be displayed through this window. The breathalyzer will include an instruction manual for the test subject to help them interpret the results and provide advice from their doctor or equivalent professional regarding possible further testing and treatment options.

[0185] Example 2 - Intake 13 After C-labeled urea was used as a marker of Helicobacter pylori infection, ammonia and... 13 Simultaneous calculation of CO2 increase.

[0186] In this embodiment, subjects avoided the use of antibiotics, bismuth preparations, proton pump inhibitors, and sucralfate for two weeks. After the two-week period, subjects fasted and abstained from water for one hour. At the end of the one-hour fast, subjects exhaled into the mouthpiece. Exhalation lasted 2 to 10 seconds, or until a specific sound or light from the main body of the breathalyzer indicated that a sufficient amount of exhaled air had entered the main body through the mouthpiece. "Sufficient" means that the exhaled sample was sufficient to come into contact with one or more gas sensors.

[0187] At the end of exhalation and when a sufficient amount of exhaled air enters the body from the mouthpiece, it first passes through a filter (if present). At the filter, gases such as NO, CH4, N2, volatile organic compounds, O2, and moisture are blocked from reaching one or more gas sensors. The filter does not block ammonia and... 13 CO2, these gases are allowed to pass through and come into contact with one or more gas sensors.

[0188] In another embodiment, exhaled gas enters the body directly from the mouthpiece and reaches one or more gas sensors. The resistivity of the one or more gas sensors depends on the presence of ammonia and [other gases] in the exhaled gas in contact with the one or more gas sensors. 13 The resistivity changes due to the amount of CO2. By using electronic circuitry and software, the resistivity changes of one or more gas sensors are converted into the ppb of ammonia. 13 CO2 ppm. Ammonia and... (The sentence is incomplete and requires more context to translate accurately.) 13 The CO2 value is stored in the memory of the electronic circuit.

[0189] At the end of baseline exhalation, the subject ingested a known amount (e.g., 125 mg, 250 mg, or 300 mg depending on the subject's weight and age). 13C-labeled urea is available in tablet form or as a snack or beverage. Post-ingestion breath measurements are taken by exhaling through a mouthpiece over a span of 10 to 120 minutes following urea ingestion, following the same procedure as baseline breath measurements. Most typically, post-ingestion breath measurements are taken 20 minutes after ingestion of the labeled urea. Post-ingestion breath measurements must not occur earlier than 10 minutes or later than 120 minutes after ingestion of the labeled urea. Ammonia concentration values ​​are expressed in ppb and... 13 The CO2 concentration value, expressed in ppm, is stored in the device's memory.

[0190] The device will calculate ammonia and... in the following manner. 13 The final value of CO2:

[0191] Final ammonia = Ammonia after urea intake minus baseline ammonia

[0192] final 13 CO2 = after urea intake 13 CO2

[0193] The designed software will convert the final ammonia value to ppb and the final 13 CO2 values ​​are converted to ppm. In subjects testing positive for Helicobacter pylori infection, baseline ammonia levels ranged from 20 ppb to 200 ppb, and post-urea intake ammonia levels ranged from 80 ppb to 600 ppb. Because there is overlap between baseline and post-urea intake ammonia levels, the percentage change between pre-urea intake and post-urea intake ammonia levels is used as a marker of Helicobacter pylori infection. A percentage change greater than 200% between pre-urea intake and post-urea intake ammonia levels indicates a positive Helicobacter pylori infection; a percentage change less than 200% indicates a negative Helicobacter pylori infection.

[0194] In subjects who tested positive for Helicobacter pylori infection, baseline 13 CO2 concentrations are typically below 200 ppm, and after urea ingestion... 13 CO2 concentrations are typically above 200 ppm (and can range from, for example, 200 ppm to approximately 1000 ppm).

[0195] In one implementation, a positive test result will be added to a value where the percentage change in ammonia is higher than 200%. 13 The percentage change in CO2 is higher than 1000%, and this value is determined using device-specific software.

[0196] In another embodiment, the device's software converts the final result into a value recognizable by a computer program, which is then calculated using an equation that takes into account ammonia and 13 The percentage increase in CO2 between baseline exhaled breath samples and exhaled breath samples after urea ingestion. This equation can be logarithmic or statistical, and includes two variables corresponding to the tested subject (ammonia and...). 13 CO2) and two constants and one or more coefficients.

[0197] In another embodiment, a positive Helicobacter pylori result will be represented by one color (e.g., red), and a negative Helicobacter pylori result will be represented by one color (e.g., green). In yet another embodiment, a positive Helicobacter pylori result will be represented by a plus sign (+), and a negative Helicobacter pylori result will be represented by a minus sign (−).

[0198] The final results can be displayed through a window on the main body of the device. The final results (e.g., indicating positive or negative, yes / no, different colored lights, or ammonia values) and... 13 The CO2 reading will be displayed through a window on the main unit. The device will also include an instruction manual to allow subjects undergoing testing to interpret the displayed results and to provide them with recommendations for possible further examinations and treatment options by a physician or equivalent.

[0199] Example 3 - Calculation of baseline human exhalation and urea intake ( 13 The proportion of C-labeled urea in exhaled air 13 CO2 / 12 The increase in CO2, i.e., exceeding the baseline (DOB), and the host-dependent urea hydrolysis rate were calculated, and both were used as markers of Helicobacter pylori infection.

[0200] In this embodiment, the subject avoids the use of antibiotics, bismuth preparations, proton pump inhibitors, and sucralfate for two weeks. At the end of the two-week period, the subject fasts and abstains from drinking for one hour. At the end of the one-hour fast, the subject exhales through the mouthpiece. Baseline exhalation occurs within 2 to 10 seconds (or longer), or until the main body of the device emits a specific sound or light indicating that a sufficient amount of exhaled gas has entered the main body through the mouthpiece. "Sufficient" means that the exhaled breath sample is sufficient to contact one or more gas sensors. In another embodiment, the subject briefly exhales through the mouthpiece for less than 5 seconds, and this can be considered a brief expiratory pulse entering the device through the mouthpiece. In this embodiment, this brief expiratory pulse will be a sufficient amount of gas to enter the device through the mouthpiece. Multiple (e.g., more than one) brief expiratory pulses may also occur that will enter the breathalyzer through the mouthpiece.

[0201] When a sufficient amount of exhaled gas enters the device body and passes through the filter, unwanted gases (e.g., NH3, NO, CH4, N2, volatile organic compounds, and / or O2) and moisture are captured and cannot pass through the filter. The filter does not block CO2 gas, which then comes into contact with one or more gas sensors.

[0202] In another embodiment, the device does not include a filter, and a sufficient amount of exhaled air enters the device 10 through the mouthpiece within 2 to 10 seconds (or an exhaled air pulse) and reaches one or more gas sensors directly (i.e., without passing through a filter). In such an example, any gas contained therein is not blocked as the exhaled air is delivered from the mouthpiece to one or more gas sensors 50.

[0203] The resistivity of at least one sensor changes based on the amount of CO2 present in the exhaled gas that comes into contact with at least one sensor. This resistivity change is converted into CO2 ppm using electronic circuitry 60 and software. The CO2 ppm value obtained during the first exhalation is stored in the device's memory using software.

[0204] In another embodiment, the sensor's resistivity is not converted to ppm. It is retained as a current measurement resulting from the resistivity change and stored in the device's memory after each exhalation (baseline and after urea ingestion).

[0205] At the end of baseline exhalation, the subject ingested a known amount (e.g., 125 mg, 250 mg, or 300 mg) of the product in the form of a capsule or tablet, or via a small meal or beverage. 13 C-labeled urea. Ingestion 13 Within a span of 10 to 120 minutes after ingesting C-labeled urea, subjects exhaled a second time into the mouthpiece following the same procedure as baseline exhalation. Most typically, exhalation after urea ingestion occurs within the timeframe of ingestion... 13 The inhalation of C-labeled urea must be performed 20 minutes after ingestion. Exhalation must not precede the inhalation of urea. 13 C-labeled urea should be ingested no later than 10 minutes after administration. 13 It occurs 120 minutes after C-labeled urea ingestion. The CO2 ppm value is measured after exhalation following urea ingestion. 13 The ppm value of CO2 is stored in the device's memory.

[0206] The device will then calculate CO2 and 13 The final value of CO2:

[0207] Final CO2 = CO2 after urea intake minus baseline CO2

[0208] final 13CO2 = after urea intake 13 CO2 minus baseline 13 CO2

[0209] In one implementation scheme, a positive test result would be: the exhaled breath after urea ingestion. 13 CO2 / CO2 ratio relative to baseline exhaled gas sample 13 The percentage change between CO2 and CO2 is greater than a specific value, referred to as the cutoff value. Above the cutoff value, the result is positive for the presence of Helicobacter pylori infection. Below the cutoff value, the result is negative for Helicobacter pylori infection. The cutoff value is calculated during clinical examination of patients with Helicobacter pylori infection undergoing this breath test. The highest cutoff value is shown in subjects who test negative for Helicobacter pylori infection (in clinical trials). 13 The CO2 / CO2 level will serve as a cutoff value. A value above this cutoff value will indicate a positive result for Helicobacter pylori infection.

[0210] In another implementation, the final result will be calculated using an equation that considers the percentage increase in CO2 between a baseline breath sample and a breath sample taken after urea ingestion. This equation can be logarithmic or statistical, and includes a variable (CO2) corresponding to the metabolic rate of the tested subject, one or more constants, and one or more coefficients. The equation will be used to determine the range of values; values ​​above this range indicate a positive result for Helicobacter pylori infection, and values ​​below this range indicate a negative result for Helicobacter pylori infection.

[0211] In another embodiment, a positive Helicobacter pylori result will be represented by one color (e.g., red), and a negative Helicobacter pylori result will be represented by one color (e.g., green). In another embodiment, a positive Helicobacter pylori result will be represented by a plus sign (+), and a negative Helicobacter pylori result will be represented by a minus sign (−).

[0212] 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...) 13 The CO2 / CO2 DOB (Difference of Biscalar Scale) value will be displayed through a window on the main body of the device. The device will also include an instruction manual to allow subjects undergoing the test to interpret the displayed results and to provide subjects with advice from a physician or equivalent professional on possible further examinations and treatment options.

[0213] Calculate the host-dependent urea hydrolysis rate (UHR):

[0214] UHR = CO2 produced × Overshoot × 0.3463, where the overshoot is defined as baseline CO2 (or13 CO2) and CO2 after urea ingestion (or 13 The difference between CO2 and CO2.

[0215] In some embodiments, this disclosure provides a multi-sensor breath analyzer (breath analyzer) device that analyzes the presence of gases in an individual's exhaled breath, including but not limited to acetaldehyde, 2-propanol, acetonitrile, acrylonitrile, benzene, isoprene, pentane, methylhexane, ethane, hydrogen sulfide, triethylamine, trimethylamine, 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), and carbon dioxide (CO2 or... 13 CO2), carbon monoxide (CO), 2,2,4,6,6-pentamethylheptane, 3,6-dimethyldecane, dodecane, 2,3,4-trimethylhexane, 2,6,8-trimethyldecane, tridecane, undecane, tetradecane, and other gases.

[0216] In one embodiment, as shown in the figure, for example, a breathalyzer device is provided, comprising a body that may be made of a durable and lightweight material. The body may include a display (e.g., a touchscreen display). The display may extend (and cover) a portion or all of one side of the body (e.g., using unbreakable glass). The display may allow user input and output after the device has completed analyzing the user's breath.

[0217] The main body of the breathalyzer ( Figure 21 This device can include multiple sensors, each independently connected to electronic circuitry and a microprocessor. Electronic circuitry, mechanically connected to each sensor, converts the sensor's resistivity and current into voltage. The microprocessor (which can be an Arduino or another type of microprocessor constructed for use in this breathalyzer) analyzes the signals from the sensors and converts them back into voltage. Through software, the voltage is converted into measurement units for each gas that the individual sensors are sensitive to.

[0218] In one embodiment, the sensor of the breathalyzer is an electrochemical sensor. The electrochemical sensor has a conductive portion and a substrate portion. The conductive portion may include, but is not limited to, gold, platinum, palladium, or a mixture of gold and platinum. The substrate portion may include, but is not limited to, a doped polymer, such as doped polyaniline, doped polypyrrole, etc. When polyaniline is doped with a protic acid (such as dinaphthalenesulfonic acid (DNNSA), camphorsulfonic acid (CSA), hydrochloric acid (HCl), salicylic acid (SSA), and 4-dodecylbenzenesulfonic acid (DBSA)), it exhibits high sensitivity to gases such as ammonia, hydrogen, nitrogen, methane, and hydrogen sulfate. Polypyrrole doped with FeCl3 or aminobenzenesulfonic acid (ABSA) or salicylic acid or another protic acid is sensitive to CO2 and... 13 CO2 exhibits sensitivity. Electrochemical sensors, such as polymer-based sensors (e.g., doped polyaniline and polypyrrole), are more effective and desirable for breath analyzers because they are stable and can operate at room temperature (around 70 degrees Fahrenheit), which is the optimal operating temperature for this breath analyzer.

[0219] In another embodiment, the breathalyzer sensor is a metal oxide nanosensor, such as ZnO, PbO-doped SnO2, which is sensitive to hydrogen, methanol, propanol, and acetone. Other metal oxides can be used as sensor materials, either alone or in combination with other conductive materials placed on metal finger electrodes (such as platinum, gold, or both), or used in floating-gate field-effect transistors (FGFETs) or carbon nanotubes or carbon nanowires.

[0220] In another embodiment, the sensor of the breathalyzer is a polymer (e.g., polyaniline and / or polypyrrole) on a conductive material, which is a chemically sensitive field-effect transistor or a floating-gate field-effect transistor (FGFET) or any other field-effect transistor (FET).

[0221] In one embodiment, the breathalyzer includes a removable mouthpiece or an opening on one side of the breathalyzer 100 configured to allow the user's mouth to cover it. In one embodiment where the device opening is used as the mouthpiece, a desiccant may be embedded in the wall of the device itself. In another embodiment where a removable mouthpiece is present, a desiccant may be embedded in the wall of the mouthpiece. The desiccant absorbs moisture at the level desired for sensor operation. Additionally or alternatively, a filter may be positioned in front of part (or all) of the sensor to block gases that may interfere with the specific sensor's task of detecting another gas. Such a filter may be in the form of crystals or silica-like crystals.

[0222] The breathalyzer's display (which may be a touchscreen and occupy the entire side of the device, much like a smartphone screen) may include drop-down menus providing users with various testing options. These options may 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, etc. After the user selects one or more medical diagnoses, the breathalyzer, through its software, requests demographic information and characteristics or symptoms consistent with the disease being tested. Once this process is complete and meets the requirements of the pre-programmed algorithm, the user is prompted to exhale into the breathalyzer. The breathalyzer then performs a predetermined task of analyzing the breath sample and provides results and general recommendations.

[0223] In one embodiment, the present invention provides a breathalyzer device comprising and utilizing 21 or more sensors, each sensor being sensitive to a gas. These gases may include acetaldehyde, 2-propanol, acetonitrile, acrylonitrile, benzene, isoprene, pentane, methylhexane, ethane, hydrogen sulfide, triethylamine, trimethylamine, 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 or... 13 The breath test device 100 can be used to screen and monitor 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 liver disease), Helicobacter pylori infection, renal failure, and metabolic diseases (diabetes, hereditary diseases). It can also detect CO2, carbon monoxide (CO), 2,2,4,6,6-pentamethylheptane, 3,6-dimethyldecane, dodecane, 2,3,4-trimethylhexane, 2,6,8-trimethyldecane, tridecane, undecane, tetradecane, and H2O. Under this embodiment, the breath test device 100 can be used to screen and monitor 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 liver disease), Helicobacter pylori infection, renal failure, and metabolic diseases (diabetes, hereditary diseases).

[0224] In another embodiment, the present invention provides a breathalyzer comprising 21 or more sensors, and electronically utilizing all or fewer sensors upon instruction, each sensor being sensitive to a gas. These gases include, but are not limited to, acetaldehyde, 2-propanol, acetonitrile, acrylonitrile, benzene, isoprene, pentane, methylhexane, ethane, hydrogen sulfide, triethylamine, trimethylamine, 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 or... 13CO2), carbon monoxide (CO), 2,2,4,6,6-pentamethylheptane, 3,6-dimethyldecane, dodecane, 2,3,4-trimethylhexane, 2,6,8-trimethyldecane, tridecane, undecane, tetradecane.

[0225] In another embodiment, the present invention provides a breathalyzer comprising 21 or more sensors, and electronically utilizing 17 or fewer sensors upon instruction, each sensor being sensitive to a gas. These gases may include, but are not limited to: 2-propanol, acetonitrile, acrylonitrile, benzene, isoprene, pentane, methylhexane, ethane, hydrogen sulfide, triethylamine, trimethylamine, carbon disulfide, dimethyl sulfide, 1-heptene, 1-octene, 1-nonene, and 1-decene.

[0226] In another embodiment, the present invention provides a breathalyzer comprising 21 or more sensors, and electronically utilizing 7 or fewer sensors upon instruction, each sensor detecting a gas. These gases may include 2-propanol, acrylonitrile, carbon disulfide, dimethyl sulfide, ethanol, isoprene, and trimethylamine.

[0227] In another embodiment, the present invention provides a breathalyzer comprising 21 or more sensors, and electronically utilizing two or fewer sensors upon instruction, each sensor detecting a gas. In this embodiment, one or two gases are detected. These gases include, but are not limited to, hydrogen (H2), nitric oxide (NO), ammonia, acetone, CO2, or... 13 CO2. Under this implementation plan, the diseases used for screening and monitoring include, but are not limited to, celiac disease, NCGS, IBS, diabetes, Helicobacter pylori infection, asthma, and kidney failure.

[0228] In another embodiment, the present invention provides a breathalyzer device comprising one or two or more sensors, each sensor being sensitive to one or more gases.

[0229] In yet another embodiment, the present invention provides a breathalyzer device comprising seven or more sensors, each sensor being sensitive to one or more gases.

[0230] In one embodiment, the mouthpiece of the breathalyzer is an opening on one side of the breathalyzer device. In another embodiment, the mouthpiece is a separate cylindrical component removably attached to the device.

[0231] In one embodiment, the breathalyzer device includes a desiccant (e.g., in crystal form) for removing a predetermined amount of moisture from a breath sample and / or a filter for preventing one or more gases from contacting the working sensor. These crystals are located inside the device near the sensor, or on or within the wall of the mouthpiece, below the mouthpiece, or inside the device.

[0232] The following non-limiting embodiments illustrate certain methods that can be used with a breathalyzer.

[0233] Example 4

[0234] Users who wish to use this breathalyzer to check if they have celiac disease can do so using the following procedure:

[0235] 1. The user presses the power button on the front of the device, and the device responds by displaying a greeting on the screen, asking for user information (e.g., name, height, weight, age, gender), and guiding the user to click the drop-down menu.

[0236] 2. The user selects (for example, clicks) the drop-down menu and finds the celiac disease / SIBO icon.

[0237] 3. The user first selects (for example, clicks) the celiac disease / SIBO icon, and then selects the celiac disease icon.

[0238] 4. The device responds through a procedure that the user follows to prepare for testing.

[0239] A) The device indicates a ready state:

[0240] B) While fasting, avoid consuming large amounts of carbohydrates for 48 consecutive hours and do not eat any food for 8 hours overnight; then exhale once into the device; and brush your teeth, but do not use mouthwash.

[0241] 5. When the user is ready, turn on the device, select (e.g., click) the drop-down menu, and select (e.g., click) "Celiac disease".

[0242] 6. The device asks the user if they are ready.

[0243] 7. The user selects (for example, by clicking) Yes (or No if not ready).

[0244] 8. The device requires the user to exhale through the mouthpiece for about 5 seconds and then wait.

[0245] 9. The user exhales for 5 seconds.

[0246] 10. Through software programming, the device uses a hydrogen sensor to detect celiac disease, shielding all other sensors and operating as a single-sensor device.

[0247] 11. The device returns a quantitative result of the amount of hydrogen in the user's breath sample and assesses whether the value is within the normal limits. The device prompts the user to discuss the test results with their healthcare provider.

[0248] Example 5:

[0249] Users wish to obtain their full metabolomics profile and can achieve this by performing the following steps. In this case, the breath analyzer utilizes all sensors simultaneously:

[0250] 1. The user turns on the device by pressing the power button.

[0251] 2. The device sends a greeting and prompts the user to select (e.g., click) a drop-down menu.

[0252] 3. From the menu, the user selects (for example, clicks) "Overview".

[0253] 4. The device responds in the following manner for the user to follow:

[0254] A) Ask for information and whether the user has used antibiotics or other types of medications.

[0255] B) Based on the response, the device requires the user to wait two weeks, or suggests fasting overnight (8 hours) and returning the following morning for testing.

[0256] 5. When users are ready in the morning, they need to brush their teeth but should not use mouthwash.

[0257] 6. The user turns on the device.

[0258] 7. The user selects (e.g., clicks) the menu.

[0259] 8. The user selects (for example, by clicking) "Overview"

[0260] 9. The device requires the user to exhale for 5 seconds through the mouthpiece and then wait.

[0261] 10. The user exhales for 5 seconds.

[0262] 11. The device utilizes all sensors simultaneously and displays readings for all detected gases, assessing whether the values ​​are within normal limits. The device prompts the user to discuss the test results with their healthcare provider.

[0263] Example 6:

[0264] Users who wish to check if they have symptoms consistent with inflammatory bowel disease (IBD) can do so by performing the following steps:

[0265] 1. Users turn on the breathalyzer by pressing the power button.

[0266] 2. The device will greet the user and ask them to select (e.g., click) a drop-down menu.

[0267] 3. The user selects IBD from the menu.

[0268] 4. The device responds in the following manner:

[0269] A) Ask for information and whether the user has used antibiotics or other medications to treat IBD;

[0270] B) Based on the answer, the device returns to the corresponding procedure.

[0271] 5. The user brushed their teeth in the morning but did not use mouthwash after fasting overnight (8 hours).

[0272] 6. The user turns on the device.

[0273] 7. The user selects (e.g., clicks) the menu.

[0274] 8. The user selects (for example, clicks) IBD.

[0275] 9. The device asks the user if they are ready.

[0276] 10. The user answered yes.

[0277] 11. The device requires the user to exhale for 5 seconds and wait.

[0278] 12. The user exhales for 5 seconds.

[0279] 13. The device shields all sensors except those that are software-programmed and operate upon IBD request.

[0280] 14. The device responds by measuring each of these gases and assessing whether it is within normal limits. The device prompts the user to discuss the test results with their healthcare provider.

[0281] Example 7:

[0282] The user suffers from asthma with inflammatory components and needs to monitor the effectiveness of treatment, and wishes to prevent relapse. In this case, the user seeks to measure nitrogen oxides in their exhaled breath, which can be achieved by performing the following steps:

[0283] 1. The user activates the breathalyzer device 100 by pressing the power button.

[0284] 2. The device provides a greeting and prompts the user to select (e.g., click) a drop-down menu icon.

[0285] 3. In the menu, the user selects (for example, clicks) Asthma / Nitric Oxide.

[0286] 4. The device returns instructions on how to perform a breath test. In this case, exhale into the device for approximately 5 to 10 seconds through the mouthpiece or opening, and ask the user if they are ready.

[0287] 5. The user selects (e.g., clicks) Yes.

[0288] 6. The device requires input data such as name, age, gender, height, and weight.

[0289] 7. After input, the device disables all sensors except for the sensor that detects nitric oxide.

[0290] 8. The device measures nitrogen oxides in the user's exhaled breath.

[0291] 9. The device stores the results and compares them with previous results from the same user.

[0292] 10. The device outputs the results on the screen and provides a trend line from previous results of tests over the past 1 to 365 days, and provides an assessment indicating whether the results are within normal limits.

[0293] 11. The device prompts the user to discuss the test results with their healthcare provider.

[0294] Although some preferred embodiments of the invention have been described, it should be understood that various modifications, adjustments and improvements can be made to these embodiments without departing from the spirit of the invention and the scope of the appended claims.

Claims

1. A self-contained, independent, handheld, battery-powered breathalyzer device that receives exhaled gas through a replaceable mouthpiece and includes: a. One or more sensors, wherein the sensors are in contact with the exhaled gas, and wherein the sensors comprise chemical and conductive materials; The chemical material is a polymer, and the resistivity of the polymer increases in response to an increase in the amount of gas in the exhaled gas sample; wherein the chemical material is in contact with the conductive material. b. A microprocessor wherein the retrieved data is fitted to a mathematical model coded in the microcontroller to generate multiple key parameters, wherein the key parameters, together with user input information, are processed by a trained machine learning model or other classification model generated from patient data and embedded in the microcontroller, and return a classification result; c. A circuit, wherein the circuit operatively connects the sensor to the microprocessor, wherein the microprocessor detects the resistivity in the circuit and uses the resistivity to calculate the concentration of gas in the exhaled gas sample.

2. The breath tester according to claim 1, wherein the sensor has a resistivity that increases with the increase of hydrogen in the breath sample.

3. The breath tester according to claim 1, wherein the sensor has a resistivity that increases with the increase of ammonia in the breath sample.

4. The breath tester according to claim 1, wherein the sensor has the ability to detect CO2 and CO2 in the breath sample. 13 The resistivity increases with the increase of CO2.

5. The breath tester according to claim 1, wherein the sensor has a resistivity that increases with the increase of methane in the breath sample.

6. The breath tester according to claim 1, wherein the sensor has a resistivity that increases with the increase of ethanol in the breath sample.

7. The breath tester according to claim 1, wherein the sensor has a resistivity that increases with the increase of acetone in the breath sample.

8. The breathalyzer according to claim 1, wherein the replaceable mouthpiece is securely attached to the body of the breathalyzer.

9. The breath tester according to claim 1, wherein the sensor has a resistivity that increases with the amount of acetaldehyde, 2-propanol, acetonitrile, acrylonitrile, benzene, isoprene, pentane, methylhexane, ethane, hydrogen sulfide, triethylamine, trimethylamine, carbon disulfide, dimethyl sulfide, 1-heptene, 1-octene, 1-nonene, 1-decene, octane, nonene, dodecane, cyclohexane, 2-butene, indole, ester, carbon disulfide, pentane, nitric oxide (NO), ethane, and propane in the breath sample.

10. The breathalyzer according to claim 1, wherein the polymer is doped polyaniline.

11. The breathalyzer according to claim 1, wherein the polymer is doped polypyrrole.

12. The breathalyzer according to claim 1, wherein the polymer is sulfonated polyaniline.

13. The breathalyzer according to claim 1, wherein the conductive material comprises a plurality of electrodes.

14. The breath tester according to claim 13, wherein the plurality of electrodes are finger electrodes and the finger electrodes are separated by a certain distance.

15. The breathalyzer of claim 1, wherein the trained file catalogs the exhaled gas based on the resistivity of the sensor, the sensor being in contact with the exhaled gas.

16. The breathalyzer of claim 15, wherein the microprocessor and the integrated circuit predict the presence of disease in a person whose exhaled gas comes into contact with the sensor.

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