Non-invasive method for diagnosing intestinal disorders

Abstract

The present invention relates to a non-invasive method for the diagnosis, prediction, risk stratification and treatment management of intestinal disorders in patients, wherein gases formed by intestinal bacteria in the exhaled gas are assayed; in particular, it relates to a pretest and the stratification of a patient.

Description

[0001] Non-invasive procedure for diagnosing bowel diseases

[0002] The present invention relates to a non-invasive method for the diagnosis, prediction, risk stratification and therapy control of intestinal diseases in patients, wherein gases produced by intestinal bacteria in the exhaled gas are determined, in particular a pre-test and the stratification of a patient.

[0003] Small intestinal bacterial overgrowth (SIBO) is an overgrowth of bacteria in the small intestine that can manifest with various symptoms such as bloating, abdominal pain, constipation, and chronic diarrhea (overview in (1)). Its prevalence is high, ranging from 2.5% to 22% of the population, depending on the study (2-4). The symptoms of SIBO and the even more common irritable bowel syndrome (IBS) often overlap (5), making SIBO testing important for accurate differential diagnosis and treatment. Conversely, misdiagnoses frequently occur, hindering treatment. Therefore, SIBO testing is also essential when IBS is suspected. Several treatment options are available for SIBO (6), primarily the administration of antibiotics / phytobiotics alongside dietary measures and probiotic supplementation.

[0004] The exact cause of SIBO remains unclear to this day, however, factors such as intestinal motility disorders, anatomical abnormalities in the intestine, certain diseases such as Crohn's disease or diabetes, as well as previous intestinal surgeries, among others, can increase the risk.

[0005] Current methods for diagnosing SIBO involve a combination of clinical examination and breath tests. These tests measure the presence of elevated levels of certain gases, such as hydrogen or methane, in the breath after ingestion of a carbohydrate solution (1, 7). These gases are metabolic products of SIBO-causing bacteria produced by the addition of sugars like glucose, lactose, or lactulose (7) and therefore allow for the specific indirect detection of small intestinal bacterial overgrowth (SIBO). Some tests rely solely on the measurement of H2, which, however, has lower sensitivity and specificity compared to a test that measures both gases (8). Furthermore, the simultaneous measurement of CO2 in breath samples is recommended to estimate non-alveolar dilution of the breath sample during collection (8, 9). The established SIBO test requires extensive preparation of the subject, which may include specific dietary measures.This includes a possible adjustment of medication. After preparation, the subject typically ingests a defined amount of a sugar (supra) and donates several more at intervals.

[0006] Breath samples are then analyzed for SIBO-typical exhaled gases. The measurement results are reviewed over time. According to guidelines from the USA and Europe, a SIBO breath test is considered positive if there is an increase of 20 ppm H2 from the baseline (8, 10) or if the absolute concentration reaches 10 ppm CH4 (8) after sugar administration.

[0007] In clinical practice, the determination of H2, CH4 and CO2 from exhaled samples is performed either by gas chromatography or gas sensors. Appropriate tests are commercially available and described in the literature (13-18).

[0008] Extensive preparation is required before performing the established tests for the detection of SIBO. For example, antibiotics and probiotics should not be taken for weeks before the test, high-fiber foods such as vegetables, especially beans and cereals, as well as certain medications should be avoided for days before the test, and the oral cavity should be disinfected with chlorhexidine on the day of the test ( 8 , 10 , 19 ).

[0009] The elaborate preparation reduces the test's acceptance, and errors during preparation also affect the test's reliability. Therefore, for screening individuals for intestinal diseases, particularly severe IBO or irritable bowel syndrome, a simple and easy-to-perform preliminary test should be required before the actual S IBO breath test. This would ensure that only those individuals with a certain probability of having intestinal diseases, especially S IBO or irritable bowel syndrome, undergo the extensive preparation.

[0010] The breath test is a non-invasive method for investigating "Small Intestinal Bacterial Overgrowth" (abbreviated: SIBO) and proceeds as follows.

[0011] Preparation: In the days leading up to the SIBO test, the test subject will be asked to avoid certain foods and medications that could affect the test result. This diet (also listed below) includes antibiotics, proton pump inhibitors, and foods high in fiber (vegetables, fruits, whole grains, legumes, nuts), especially those with more than 4% or more than 6% fiber by weight.

[0012] Test substance: The subject receives a test substance, which is usually lactulose or glucose. These substances are primarily carbohydrates that are not normally digested in the small intestine but can be fermented by intestinal bacteria, provoking gas production.

[0013] Breath sampling: The subject breathes into special breathing bags or tubes that can absorb exhaled gases. The first breath sample is taken before ingestion of the test substance to determine the baseline value. The subject then ingests the test substance.

[0014] Measurements / Determinations: After the test substance has been ingested, breath samples are taken at regular intervals, usually every 15-30 minutes over a period of several hours. These samples are then analyzed for their content of hydrogen (H2) and / or methane (CH4), which are produced by the intestinal bacteria when they ferment the test substance.

[0015] Interpretation of the results: An increase in hydrogen and / or methane levels in the breath indicates bacterial overgrowth in the small intestine. The patterns of hydrogen and methane production can also help to differentiate between various types of SIBO.

[0016] Surprisingly, it has now been found that a step test to determine the levels of the gases H2 and CH4 in the exhalation of a subject at two time points and at a time interval after a short-term fast, namely a base sample followed by a meal and a subsequent second sample, facilitates the success of a subsequent SIBO test or increases the diagnostic value of an SIBO test.

[0017] Therefore, one object of the present invention is to provide a pre-test so that patients can be stratified for further diagnosis or prediction of intestinal diseases.

[0018] Therefore, the invention relates to a non-invasive method for diagnosing or predicting intestinal diseases, wherein gases produced by intestinal bacteria are determined in the exhaled gas of a subject, comprising the following steps: a) Performing a short-term fast of at least 6 hours and / or a maximum of 12 hours, b) Determining the ppm concentrations of H2 and CH4 in the exhaled gas of a subject from a) or of a fasting subject, c) Ingestion of a meal by a subject from b), d) Determining the ppm concentrations of H2 and CH4 in the exhaled gas of a subject from c) or of the same non-fasting subject, dl) wherein an increase in the concentrations of at least more than 10 ppm of H2 and of at least more than 10 ppm of CH4 compared to b) is indicative of an intestinal disease, and the subject is stratified, e)) to verify the diagnosis or prediction of intestinal diseases by a second non-invasive procedure for the diagnosis or prediction of intestinal diseases, wherein H2 and / or CH4 are determined in the exhaled gas of a subject, or d.2) wherein an increase in the levels of H2 and CH4 of 10 ppm or less than 10 ppm or less than b.) is not indicative of SIBO, and the subject is stratified, f.) not to verify the diagnosis or prediction of intestinal diseases by a second non-invasive procedure for the diagnosis or prediction of intestinal diseases, in particular SIBO, wherein H2 and / or CH4 are determined in the exhaled gas of a subject.

[0019] The invention also relates to the use of the gases H2 and CH4, preferably in a non-invasive method, for the diagnosis or prediction of intestinal diseases, wherein gases produced by intestinal bacteria are determined in the exhaled gas of a subject, comprising the following steps: a.) performing a short-term fast of at least 6 hours and / or a maximum of 12 hours, b.) determining the ppm concentrations of H2 and CH4 in the exhaled gas of a subject from a.) or of a fasting subject, c.) ingesting a meal by a subject from b.), d.) determining the ppm concentrations of H2 and CH4 in the exhaled gas of a subject from c.) or of the same non-fasting subject, dl.) wherein an increase in the concentrations of at least more than 10 ppm of H2 and of at least more than 10 ppm of CH4 compared to b.) is indicative of an intestinal disease, and the subject is stratified, e.) to verify the diagnosis or prediction of intestinal diseases by a second use of gases H2 and CH4, preferably in a non-invasive procedure for the diagnosis or prediction of intestinal diseases, wherein H2 and / or CH4 are determined in the exhaled gas of a subject, or d.2) wherein an increase in the levels of H2 equal to or less than 10 ppm and of CH4 equal to or less than 10 ppm compared to b.) is not indicative of SIBO, and the subject is stratified, f.) to verify the diagnosis or prediction of intestinal diseases not by a second use of gases H2 and CH4, preferably in a non-invasive procedure for the diagnosis or prediction of intestinal diseases, in particular SIBO, wherein H2 and / or CH4 are determined in the exhaled gas of a subject.

[0020] Furthermore, in a further preferred embodiment, the invention relates to a non-invasive method for the diagnosis or prediction of intestinal diseases, or the use of gases H2 and CH4, preferably in a non-invasive method as above, wherein step e.) comprises the following further steps: i.) Carrying out a diet of at least 3 days of a

[0021] Subjects, ii.) Determination of the ppm levels of H2 and CH4 in the

[0022] Exhaled gas of a subject from i.) , iii.) intake of sugar, especially lactulose or

[0023] Glucose of a subject from ii.) , iv.) Determination of the ppm levels of H2 and / or CH4 in the exhaled gas of a subject from iii.) .

[0024] Furthermore, in a further preferred embodiment, the invention relates to a non-invasive method for the diagnosis or prediction of intestinal diseases, or the use of gases H2 and CH4, preferably in a non-invasive method as above, wherein step e.) , comprises the following further step: v.) wherein an increase in the levels of at least more than 10 ppm of H2 and / or of at least more than 10 ppm of CH4 compared to ii.) is indicative of SIBO.

[0025] Furthermore, in a further preferred embodiment, the invention relates to a non-invasive method for the diagnosis or prediction of intestinal diseases, or the use of gases H2 and CH4, preferably in a non-invasive method as above, wherein the additional alternative d.3) to dl) or d.2) follows: or d.3) wherein an increase in the levels of H2 equal to or less than 10 ppm and CH4 equal to or less than 10 ppm compared to b.) is not indicative of SIBO, and the subject is stratified, f.) the diagnosis or prediction of intestinal diseases by a further method for the diagnosis or prediction of at least one intestinal disease selected from the group consisting of irritable bowel syndrome (IBS), lactose or fructose malabsorption, chronic inflammatory bowel diseases (GED), in particular Crohn's disease and ulcerative colitis, celiac disease, and functional dyspepsia.

[0026] The intestinal diseases “irritable bowel syndrome (IBS), lactose or fructose malabsorption, chronic inflammatory bowel diseases (IBD), especially Crohn’s disease and ulcerative colitis, celiac disease, functional dyspepsia” exhibit similar symptoms to SIBO. These intestinal diseases are described, for example, in Pschyrembel’s medical dictionary or are known to the WHO (ICD), and suitable methods for diagnosis or prediction are known to specialists.

[0027] For the purposes of this invention, intestinal diseases such as SIBO, with SIBO being particularly preferred, are included, but also intestinal diseases that are similar to SIBO in their symptoms, namely irritable bowel syndrome (IBS), lactose or fructose malabsorption, chronic inflammatory bowel diseases (IBD), in particular Crohn's disease and ulcerative colitis, celiac disease, or functional dyspepsia. In particular, intestinal diseases exhibiting malabsorption are included according to the invention.

[0028] Furthermore, in a further preferred embodiment, the invention relates to a non-invasive method for the diagnosis or prediction of intestinal diseases, or the use of gases H2 and CH4, preferably in a non-invasive method as above, wherein an increase in the ppm content of H2 and / or CH4 takes place within 60 to 100 minutes, in particular 80 minutes.

[0029] Another function of the invention is also to

[0030] Therapy control using a prediction or diagnosis of

[0031] Intestinal diseases, preferably S IBO, wherein a subject is examined according to a method according to the invention, with the provision that the subject receives medication, in particular antibiotics / phytobiotics for the treatment of an intestinal disease or adheres to a diet for the treatment of an intestinal disease, in particular for the treatment of malabsorption, so that a therapeutic success or improvement of therapy can be achieved.

[0032] The term "fasting (period)" as used in this invention refers to a planned period during which people consciously abstain from all food and drink except for small amounts of water. The term "fasting (period)" can be used synonymously.

[0033] The term "meal" as used in this invention refers to a planned occasion in which people consume food to satisfy their hunger and provide their bodies with nutrients. A meal may, in particular, contain carbohydrates.

[0034] The term "short-term fasting of at least 6 hours and / or a maximum of 12 hours" as used in this invention means that a specific time window is defined for eating and fasting. During this fasting period, no food is consumed, in particular by means of a meal. The term "fasting" can be used synonymously.

[0035] “Diagnosis” or “prediction” within the meaning of this invention means the positive determination or prediction / prognosis or probability of the onset and occurrence of an intestinal disease, in particular SIBO.

[0036] The term diagnosis also includes medical diagnostics and related examinations, in particular in-vitro diagnostics and laboratory diagnostics.

[0037] The diagnosis according to the invention takes place outside (ex vivo) of the human body, since the respiratory gases, or rather the determination of the respiratory gases according to the invention, is carried out using gas chromatography or gas sensors. Furthermore, the presence of a physician is not required.

[0038] Furthermore, the invention relates to a method for stratification, in particular for risk stratification and / or therapy control of a patient.

[0039] “Stratification or therapy control” within the meaning of this invention means that the inventive method or the inventive use of gases H2 and CH4 allows decisions regarding the treatment and therapy of the subject, be it hospitalization of the patient, use, effect and / or dosage of one or more drugs, a therapeutic measure, or the monitoring of a disease course as well as the course of therapy or the etiology or classification of intestinal diseases, in particular SIBO. Furthermore, “stratification” within the meaning of this invention means the claimed instruction that the subject receives or does not receive a further method for the diagnosis and prediction of intestinal diseases (supra).

[0040] In another embodiment of the invention, the term “stratification” includes in particular risk stratification with the prediction of an “outcome” of an adverse health event, in particular that an intestinal disease is present.

[0041] Within the scope of this invention, "subject" is understood to mean any human being or mammal, in particular a patient. Patients who already exhibit symptoms such as bloating, abdominal pain, diarrhea, constipation, nutrient deficiencies, and weight loss due to malabsorption are particularly preferred. The invention will now be described in more detail in the following examples, without being limited thereto.

[0042] Examples:

[0043] Example 1:

[0044] For the spot test, samples are taken using plastic straws from Medi-Inn (Hirten, Germany), article number: 93652-Karton, EAN: 4260655961231 and glass tubes from Zhejiang ALWSCI Technologies Co., Ltd.

[0045] The measurements are performed using gas chromatography with a barrier discharge detector (GC-BID), equipped with a Carboxen PLOT column (30 mm x 0.53 mm and 0.50 pm layer thickness) and an ACC-6000 Plus autosampler (Shimadzu, Duisburg, Germany). The temperature gradient is 35°C (holding time 3 min) and is ramped up to 80°C (50°C / min, holding time 5 min) and further to 170°C (50°C / min, holding time 2 min). The BID and injection temperatures are maintained at 250°C. The injection volume and carrier gas flow of helium (purity 99.9999%) are 100 pL and 8 mL / min, respectively.

[0046] To collect the sample, the subject exhales through the straw; during the last third of an exhalation cycle, the lower end of the straw is inserted into a glass tube and positioned near the bottom, while exhalation continues.

[0047] The straw is then slowly moved towards the opening of the glass tube while exhaling continuously. Once the straw reaches the top of the glass tube at the end of the exhalation cycle, the tube is sealed with a screw cap. Samples for comparative measurements and the main test are taken using the EasySampler® Kit from QuinTron (3712 West Pierce Street, Milwaukee, WI, USA) according to the manufacturer's instructions. Preparation for the main and comparative tests is carried out in accordance with the guidelines for S IBO diagnostics and as described in the package insert for the VOC-S IBO test from VOC-Advanced Breath Diagnostics GmbH (Henkestraße 91, 91052 Erlangen, Germany).

[0048] The samples (preliminary test and main / comparative test) were then analyzed using a Shimadzu GC-2030 gas chromatography system with the AOC-6000 Plus autosampler and an SH-Rt-Molsieve 5A P / N 221-75763-30 column. The analysis of the measurement data was performed against a calibration curve generated using gas samples of known concentration and metrological traceability to international standards (gases were obtained from All-In Gas, Emil-Riedel-Straße 1, 80538 Munich, Germany).

[0049] Table 1:

[0050] test

[0051] Interpretation SIBO Test Match

[0052] Test No. Subject H2 CH4 Total pos / neg yes / no

[0053] 1. neg pos pos pos ja

[0054] 2. negative negative negative negative yes

[0055] 3. neg pos pos pos yes

[0056] 4. negative negative negative negative yes

[0057] 5. negative negative negative negative yes

[0058] 6. neg pos pos pos ja

[0059] 7. neg neg neg neg yes

[0060] 8. neg neg neg neg yes

[0061] 9. neg pos pos pos ja

[0062] 10. neg pos pos pos ja

[0063] 11. neg neg neg neg yes

[0064] 12. neg pos pos pos ja

[0065] 1 . pos neg pos pos ja

[0066] 14. pos neg pos pos ja

[0067] 15. neg neg neg neg yes

[0068] 16. neg neg neg neg yes

[0069] 17. neg neg neg neg yes

[0070] 18. pos neg pos pos ja

[0071] 19. neg pos pos pos ja

[0072] 20. neg neg neg neg yes

[0073] 21. neg neg neg neg yes

[0074] 22. neg neg neg neg yes

[0075] 23. neg neg neg neg yes

[0076] 24. neg neg neg neg yes 25. pos pos pos pos yes

[0077] 26. neg neg neg neg yes

[0078] 27. pos pos pos pos yes

[0079] 28. no no no no yes

[0080] Table la - 29- 109 : Test Interpretation SIBO Test Match H2 CH4 Gesamt pos / neg ja / nein neg pos pos pos yes neg neg neg neg yes neg neg neg neg yes neg neg neg neg yes neg neg neg pos no neg neg neg neg yes neg neg neg neg yes neg neg neg neg yes neg neg neg neg yes neg neg neg neg yes neg neg neg yes pos neg pos pos yes neg pos pos pos yes neg neg neg neg yes neg neg neg neg yes neg neg neg neg yes neg pos pos neg yes neg pos pos pos yes neg neg neg neg yes neg neg neg neg yes neg neg neg neg yes neg neg neg yes pos neg pos pos yes neg neg neg neg yes neg neg neg yes neg neg neg yes pos neg pos pos yes neg neg neg neg yes neg neg neg yes neg neg neg neg yes neg neg neg neg yes pos neg pos pos neg neg yes neg neg neg yes neg neg neg neg yes neg neg neg yes neg neg neg neg yes neg pos pos yes neg neg neg yes neg neg neg pos no neg neg neg neg yes neg neg neg neg yes neg neg neg neg pos pos yes neg neg neg yes neg neg neg yes 84 neg neg neg yes

[0081] 85 neg pos pos pos yes

[0082] 86 neg pos pos pos yes

[0083] 87 pos neg pos pos yes

[0084] 88 neg neg neg yes

[0085] 89 negative negative negative yes

[0086] 90 negative negative negative yes

[0087] 91 negative negative negative yes

[0088] 92 negative negative negative yes

[0089] 93 neg pos pos pos yes

[0090] 94 negative negative negative yes

[0091] 95 negative negative negative yes

[0092] 96 negative negative negative yes

[0093] 97 post post post already

[0094] 98 neg neg neg yes

[0095] 99 negative negative negative yes

[0096] 100 negative negative negative yes

[0097] 101 neg neg neg yes

[0098] 102 neg pos pos pos yes

[0099] 103 pos neg pos pos yes

[0100] 104 neg pos pos pos yes

[0101] 105 negative negative negative yes

[0102] 106 negative negative negative yes

[0103] 107 negative negative negative yes

[0104] 108 negative negative negative yes

[0105] 109 negative negative negative yes

[0106] Interpretation: SPOT positive at Anstieg H2 from time point 0 mm at 80 min to confirm > 10 ppm and CH4 h 10 ppm

[0107] Time of day. Main test positive if H2 rises within 90 min after substrate addition > 20 ppm and CH4 rises above 10 ppm at any time.

[0108] Conclusion: The spot test is suitable for pre-selection of patients for the application of the full test for S IBO, but also lactose or fructose malabsorption.

[0109] Result: Prediction of the spot test at cut of f (threshold) of more than 10 ppm for hydrogen and methane is 100% correct at N=28 or even N=109.

[0110] Literature:

[0111] 1. Skrzydlo-Radomanska B, Cukrowska B. How to Recognize and Treat Small Intestinal Bacterial Overgrowth? J Clin Med. 2022 Oct 12; 11(20):6017)

[0112] 2. Krajicek, E. J.; Hansel, SL Small Intestinal Bacterial Overgrowth: A Primary Care Review. Mayo Clin. Proc. 2016, 91, 1828-1833

[0113] 3. Achufusi, T.G.O.; Sharma, A.; Zamora, E.A.; Manocha, D. Small Intestinal Bacterial Overgrowth: Comprehensive Review of Diagnosis, Prevention, and Treatment Methods. Cureus 2020, 12, e8860

[0114] 4. Grace, E.; Shaw, C.; Whelan, K.; Andreyev, H.J.N. Review article: Small intestinal bacterial overgrowth-Prevalence, clinical features, current and developing diagnostic tests, and treatment. Aliment. Pharmacol. Ther. 2013, 38, 674-688

[0115] 5. Ghoshal UC, Shukla R, Ghoshal U. Small Intestinal Bacterial Overgrowth and Irritable Bowel Syndrome: A Bridge between Functional Organic Dichotomy. Gut Liver. 2017 Mar 15;11 (2): 196-208. doi: 10.5009 / gnll6126. PMID: 28274108; PMCID: PMC5347643.

[0116] 6. Rao S.S.C., Bhagatwala J. Small Intestinal Bacterial Overgrowth: Clinical features and therapeutic management: Clinical features and therapeutic management. Clin. Transl. Gastroenterol. 2019;10:e00078.

[0117] 7. Losurdo G., Leandro G., lerardi E., Perri F., Barone M., Principi M., Di Leo A. Breath Tests for the Non-invasive Diagnosis of Small Intestinal Bacterial Overgrowth: A Systematic Review with Meta-analysis. J. Neurogastroenterol.

[0118] 8. Rezaie A, Buresi M, Lembo A, Lin H, McCallum R, Rao S, Schmulson M, Valdovinos M, Zakko S, Pimentel M. Hydrogen and Methane-Based Breath Testing in Gastrointestinal Disorders: The North American Consensus. Am J Gastroenterol. 2017 May;112(5):775-784. Goldoni M, Corradi M, Mozzoni P, Folesani G, Alinovi R, Pinelli S, Andreoli R, Pigini D, Tillo R, Filetti A, Garavelli C, Mutti A. Concentration of exhaled breath condensate biomarkers after fractionated collection based on exhaled CO2 signal. J Breath Res. 2013 Mar;7(l) Gasbarrini A, Corazza GR, Gasbarrini G, Montalto M, Di Stefano M, Basilisco G, Parodi A, Usai-Satta P, Vemia P, Anania C, Astegiano M, Barbara G, Benini L, Bonazzi P, Capurso G, Certo M, Colecchia A, Cuoco L, Di Sario A, Festi D, Lauritano C, Miceli E, Nardone G, Perri F, Portincasa P, Risicato R, Sorge M, Tursi A; 1st Rome H2-Breath Testing Consensus Conference Working Group.Methodology and indications of H2-breath testing in gastrointestinal diseases: the Rome Consensus Conference. Aliment Pharmacol Ther. 2009 Mar 30;29 Suppl 1 : 1-49. Banik GD, De A, Som S, Jana S, Daschakraborty SB, Chaudhuri S, Pradhan M. Hydrogen sulphide in exhaled breath: a potential biomarker for small intestinal bacterial overgrowth in IBS. J Breath Res. 2016 May 10;10(2):026010. doi: 10.1088 / 1752- 7155 / 10 / 2 / 026010. PMID: 27163246. Takakura W, Pimentel M. Small Intestinal Bacterial Overgrowth and Irritable Bowel Syndrome - An Update. Front Psychiatry. 2020 Jul 10; 11 :664. doi: 10.3389 / fpsyt.2020.00664. PMID: 32754068; PMCID: PMC7366247. Shrestha A, Prodhan UK, Mitchell SM, Sharma P, Barnett MPG, Milan AM, Cameron- Smith D. Validity of a Portable Breath Analyser (AIRE) for the Assessment of Lactose Malabsorption. Nutrients.2019 Jul 17; 11(7): 1636 Guillermo Barahona, Barry Me Bride, Aine Moran, Sahar Hawamdeh, Luisa Villatoro, Robert Bums, Bo Konings, Robert Bulat, Megan McKnight, Claire Shortt, Pankaj J. Pasricha;Improving the Diagnosis of SIBO Using an At-Home Handheld App Connected Breath Analysis Device (AIRE). medRxiv preprint doi: https: / / doi.org / 10.1101 / 2022.04.21.22274143; this version posted April 21, 2022. Saad RJ, Chey WD. Breath testing for small intestinal bacterial overgrowth: maximizing test accuracy. Clin Gastroenterol Hepatol. 2014 Dec; 12(12): 1964-72; Dharmawardana N, Goddard T, Woods C, Watson DI, Butler R, Ooi EH, Yazbeck R. Breath methane to hydrogen ratio as a surrogate marker of intestinal dysbiosis in head and neck cancer. Sei Rep. 2020 Sep 14; 10(1): 15010 de Lacy Costello BP, Ledochowski M, Ratcliffe NM. The importance of methane breath testing: a review. J Breath Res. 2013 Jun;7(2):024001.Gao, Fan & Wang, Min & Zhang, Xusheng & Zhang, Junyu & Xue, Yingying & Wan, Hao & Wang, Ping. (2018). Simultaneous Detection of Hydrogen and Methane in Breath for the Diagnosis of Small Intestinal Bacterial Overgrowth by Fast Gas Chromatography. Analytical Methods. 10. 10.1039 / C8AY01451E. Pique JM, Pallares M, Cuso E, Vilar-Bonet J, Gassull MA. Methane production and colon cancer. Gastroenterology. 1984 Sep;87(3):601-5. PMID: 6745612. Li L, Zhang XY, Yu JS, Zhou HM, Qin Y, Xie WR, Ding WJ, He XX. Ability of lactulose breath test results to accurately identify colorectal polyps through the measurement of small intestine bacterial overgrowth. World J Gastrointest Surg. 2023 Jun 27;15(6): 1138-1148. doi: 10.4240 / wjgs.vl5.i6.1138. PMID: 37405104; PMCID: PMC10315122. Lin H, Yu Y, Zhu L, Lai N, Zhang L, Guo Y, Lin X, Yang D, Ren N, Zhu Z, Dong Q. Implications of hydrogen sulfide in colorectal cancer: Mechanistic insights and diagnostic and therapeutic strategies. Redox Biol.2023 Feb;59: 102601. doi: 10.1016 / j. redox.2023.102601. Epub 2023 Jan 7. PMID: 36630819; PMCID: PMC9841368. Saad RJ, Chey WD. Breath testing for small intestinal bacterial overgrowth: maximizing test accuracy. Clin Gastroenterol Hepatol. 2014 Dec; 12(12): 1964-72; quiz el 19-20. Gottlieb K, Le C, Wacher V, Sliman J, Cruz C, Porter T, Carter S. Selection of a cutoff for high- and low-methane producers using a spot-methane breath test: results from a large north American dataset of hydrogen, methane and carbon dioxide measurements in breath. Gastroenterol Rep (Oxf). 2017 Aug;5(3): 193-199. doi:.

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Claims

Patent claims:

1. A non-invasive method for diagnosing or predicting intestinal diseases, wherein gases produced by intestinal bacteria in the exhaled gas of a subject are determined, comprising the following steps: a.) performing a short-term fast of at least 6 hours and at most 12 hours on a subject, b.) determining the ppm concentrations of H2 and CH4 in the exhaled gas of a subject from a.), c.) ingesting a meal by a subject from b.), d.) determining the ppm concentrations of H2 and CH4 in the exhaled gas of a subject from c.), dl.) wherein an increase in the concentrations of at least more than 10 ppm of H2 and at least more than 10 ppm of CH4 compared to b.) is indicative of an intestinal disease, and the subject is stratified, e.) verifying the diagnosis or prediction of intestinal diseases by a second non-invasive method for diagnosing or predicting intestinal diseases, wherein H2 and / or CH4 in the exhaled gas of a subject can be determined, or d.2) wherein an increase in the levels of H2 and CH4 of 10 ppm or less than 10 ppm compared to b.) is not indicative of SIBO, and the subject is stratified, f.) not to verify the diagnosis or prediction of intestinal diseases by a second non-invasive procedure for the diagnosis or prediction of intestinal diseases, in particular SIBO, whereby H2 and / or CH4 are determined in the exhaled gas of a subject.

2. A non-invasive method for diagnosing or predicting intestinal diseases according to claim 1, wherein step e.) comprises the following further steps: i.) carrying out a diet of at least 3 days of a Subjects, ii.) Determination of the ppm levels of H2 and CH4 in the Exhaled gas of a subject from i.) , iii.) intake of sugar, especially lactulose or Glucose of a subject from ii.) , iv.) Determination of the ppm levels of H2 and / or CH4 in the exhaled gas of a subject from iii.) .

3. Non-invasive method for diagnosing or predicting intestinal diseases according to claim 2, wherein step e.) , comprises the following further step: v.) wherein an increase in the levels of at least more than 10 ppm of H2 and / or of at least more than 10 ppm of CH4 compared to ii.) is indicative of SIBO.

4. A non-invasive method for diagnosing or predicting intestinal diseases according to claim 1, wherein the additional alternative d.3) follows: or d.3) wherein an increase in the levels of H2 equal to or less than 10 ppm and of CH4 equal to or less than 10 ppm compared to b.) is not indicative of SIBO, and the subject is stratified, f.) the diagnosis or prediction of intestinal diseases by a further method for diagnosing or predicting at least one intestinal disease selected from the group consisting of irritable bowel syndrome (IBS), lactose or fructose malabsorption, chronic inflammatory bowel diseases To conduct examinations for intestinal diseases (IBD), especially Crohn's disease and ulcerative colitis, celiac disease, and functional dyspepsia.

5. Non-invasive method for diagnosing or predicting intestinal diseases according to any one of claims 1 to 4, wherein an increase in the ppm levels of H2 and / or CH4 occurs within 60 to 100 minutes, in particular 80 minutes.

6. Non-invasive method for diagnosing or predicting intestinal diseases according to any one of claims 1 to 5, for therapy control, with the provision that the subject receives medication, in particular antibiotics, phytobiotics for the treatment of an intestinal disease or adheres to a diet for the treatment of an intestinal disease, in particular for the treatment of malabsorption.

7. Non-invasive method for diagnosing or predicting intestinal diseases according to any one of claims 1 to 5 for risk stratification of subjects.

8. Non-invasive method for diagnosing or predicting intestinal diseases according to any one of claims 1 to 7, wherein gases produced by intestinal bacteria are determined in the exhaled gas of a subject, wherein the determination of H2 and CH4 is carried out by means of gas chromatography or gas sensors outside the human body.

9. Use of H2 and CH4 gases produced by intestinal bacteria in the exhaled gas of a subject for the diagnosis or prediction of intestinal diseases according to any one of claims 1 to 8.

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