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Reference Guide
Gemelli Biotech
trio-smart SIBO Breath Test

trio-smart SIBO Breath Test

Gemelli Biotech
trio-smart SIBO Breath Test
Gemelli Biotech
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About the Test

The Trio-Smart SIBO breath test by Gemelli Biotech offers a comprehensive diagnostic approach for individuals experiencing gastrointestinal symptoms such as bloating, gas, abdominal pain, constipation, and diarrhea. 

By measuring hydrogen, methane, and hydrogen sulfide levels in the breath, this test can detect three variations of bacterial overgrowth: small intestinal bacterial overgrowth (SIBO), intestinal methanogenic overgrowth (IMO), and excess hydrogen sulfide. [12., 13.]

Through at-home sample collection and analysis at a CLIA-certified lab, patients and healthcare providers gain insight into the underlying causes of gastrointestinal symptoms, facilitating the development of tailored treatment plans. [12.]

Breath tests are effective in detecting bacterial overgrowth conditions like small intestinal bacterial overgrowth (SIBO), which are strongly linked to GI symptoms.  These tests offer a noninvasive and cost-effective approach to diagnosing GI disorders, providing valuable insights for effective management.

Many individuals with functional gut disorders may not realize the impact of their diet on their GI symptoms. Particularly, those experiencing chronic symptoms like abdominal pain, bloating, flatulence, and altered bowel movements may overlook the possibility of carbohydrate malabsorption or SIBO. Breath tests offer specific and sensitive diagnostic capabilities, allowing healthcare providers to confirm or rule out these conditions in such patients.

Elevated levels of hydrogen, methane, and hydrogen sulfide can indicate disruptions in the microbiome, leading to various gastrointestinal symptoms. Depending on the substrate used, a carbohydrate malabsorption may also be discovered.  

The Trio-Smart test provides data to guide treatment decisions, offering patients a personalized approach to addressing their symptoms and improving their overall well-being.

What is the Science Behind the trio-smart SIBO Breath Test By Gemelli Biotech?

Breath testing serves as a valuable tool in identifying various gut microbiome-related conditions by analyzing fermented gasses in exhaled breath samples. The three primary gasses measured in breath testing are hydrogen, methane, and hydrogen sulfide, which can provide insights into disorders affecting gut health. These gasses are produced by bacteria in the gastrointestinal tract during the fermentation of sugars,  [11.]

High levels of hydrogen, methane, or hydrogen sulfide in breath testing can have distinct clinical implications. 

Elevated hydrogen levels are commonly associated with small intestinal bacterial overgrowth (SIBO), where excess bacteria in the small intestine produce hydrogen gas during the fermentation of carbohydrates. This can lead to symptoms such as bloating, abdominal discomfort, and diarrhea.  [13.]

High methane levels are indicative of intestinal methanogenic overgrowth (IMO), which is often linked to constipation. Methane is produced by archaea in the gut that consume hydrogen, slowing gut transit time and potentially contributing to constipation.  [13.]

In contrast, elevated hydrogen sulfide levels in breath testing are associated with excess hydrogen sulfide production in the gut microbiome. This condition, previously undetectable by traditional breath tests, is linked to symptoms such as diarrhea and abdominal pain. Excess hydrogen sulfide is believed to be produced by sulfate-reducing bacteria and can lead to gut inflammation and dysbiosis.

Each gas—hydrogen, methane, and hydrogen sulfide—provides valuable clinical information about the underlying gut microbiome composition and function, helping healthcare providers tailor treatment strategies for patients with gastrointestinal symptoms. Therefore, understanding the clinical relevance of high levels of these gasses in breath testing is crucial for accurate diagnosis and management of gut-related disorders.

Additionally, the substrate chosen may also provide clinical insight.  While glucose hydrogen breath tests are preferred for diagnosing SIBO, lactose and fructose hydrogen breath tests are utilized to identify lactose and fructose malabsorption, respectively.

What is the Clinical Relevance of the trio-smart SIBO Breath Test By Gemelli Biotech?

A positive finding on the trio-smart SIBO Breath Test indicates microbial overgrowth in the small intestine, which can lead to gastrointestinal symptoms such as bloating, abdominal pain, gas, and diarrhea. [8.]  This test offers a non-invasive and efficient method for identifying microbial overgrowth, allowing healthcare providers to make informed dietary recommendations and treatment decisions.

A positive result on the Trio-Smart SIBO breath test indicates the presence of elevated levels of hydrogen and/or methane gasses in the breath after consuming a substrate (often lactulose or glucose). These gasses are produced by bacteria fermenting the substrate in the small intestine, which is characteristic of SIBO. [14.]

Glucose is easily absorbed in the proximal small intestine, making it an optimal substrate to assess for SIBO.  Lactulose passes intact into the colon, making it an ideal substrate to assess bowel transit time.  [5., 14.]

The clinical relevance of diagnosing and treating SIBO lies in alleviating symptoms and improving overall gastrointestinal health.  It is important to consider the likely pathogenesis of SIBO, which often involves intestinal dysmotility and decreased gastric acid production.  [3.]

By identifying SIBO through tests like the trio-smart SIBO breath test, healthcare providers can tailor treatment strategies such as antibiotics or antimicrobials, dietary modifications, lifestyle interventions including stress reduction and/or vagal nerve stimulation, and consider the judicious use of probiotics to target the overgrowth of bacteria in the small intestine.

When Should Clinicians Order the Following Substrates: Fructose, Lactose, or Sucrose?

Fructose, lactose, and sucrose are other commonly used substrates in breath testing for bacterial overgrowth as well as for malabsorption.  The choice of substrate depends on the suspected underlying cause of the patient's symptoms and their dietary habits. [11.]

Fructose is often used to assess for fructose malabsorption, which can contribute to symptoms of bloating, gas, and diarrhea. Patients suspected of having fructose intolerance may undergo breath testing with fructose to evaluate their ability to absorb this sugar. 

Lactose, found in dairy products, is commonly used to assess for lactose intolerance. Breath testing with lactose can help diagnose lactase deficiency, which leads to the inability to digest lactose and can result in symptoms such as bloating, abdominal pain, and diarrhea after consuming dairy products.

Sucrose, although less commonly used compared to fructose and lactose, can also be used as a substrate in breath testing for SIBO. Sucrose is a disaccharide composed of glucose and fructose, and its malabsorption may contribute to gastrointestinal symptoms in certain individuals. [6.]

Lactulose is an additional substrate used for breath testing.  Lactulose hydrogen breath tests are commonly used to assess GI motility by measuring orocecal transit time. [11.]

Breath testing with sucrose can help identify patients with sucrase-isomaltase deficiency or other conditions affecting the digestion and absorption of sucrose. Overall, the selection of substrate for SIBO breath testing should be based on the patient's clinical presentation, dietary history, and suspected underlying disorders, with the goal of accurately diagnosing and managing their gastrointestinal symptoms.

Who Can Benefit from the trio-smart Malabsorption Breath Test- Lactose?

Individuals with Unexplained Gastrointestinal Symptoms: Those experiencing chronic symptoms such as bloating, gas, abdominal pain, and altered bowel habits (diarrhea or constipation) could benefit from the trio-smart malabsorption breath test for lactose. Studies have shown that lactose malabsorption can contribute to these symptoms.  [1.]

Patients with Suspected Irritable Bowel Syndrome (IBS): IBS is a common condition characterized by abdominal discomfort or pain and altered bowel habits. Individuals with suspected IBS who have not responded to conventional treatments may benefit from testing for lactose malabsorption. [2.]

Individuals with Suspected Lactose Intolerance: lactose intolerance is a condition where the body has difficulty digesting and absorbing lactose due to a genetic deficiency of the enzyme lactase, leading to gastrointestinal discomfort. The trio-smart malabsorption breath test with lactose can help diagnose lactose intolerance and guide dietary management.  [1.]

Patients with Unexplained Abdominal Discomfort: Those experiencing unexplained abdominal discomfort or pain, especially after consuming foods high in lactose, may benefit from lactose breath testing. Research suggests that identifying and addressing lactose malabsorption can alleviate symptoms and improve quality of life [1., 8.]

Individuals Seeking Dietary Management: For individuals looking to optimize their diet and improve gastrointestinal symptoms, identifying lactose malabsorption through breath testing can guide dietary modifications. This personalized approach can help individuals avoid foods high in lactose and alleviate symptoms associated with malabsorption, possibly by supplementing with the enzyme lactase.  [7.]


Results should be released to your provider 7 business days after samples are received at the lab. Rupa Health cannot guarantee processing time.

Additional Resources for Clinicians

Article: SIBO Testing 101: A Complete Guide to The Top 3 SIBO Tests 

Article: How to Use the trio-smart Test from Gemelli Biotech in Clinic


[1.] Bouchoucha M, Fysekidis M, Rompteaux P, Raynaud JJ, Sabate JM, Benamouzig R. Lactose Sensitivity and Lactose Malabsorption: The 2 Faces of Lactose Intolerance. Journal of Neurogastroenterology and Motility. 2021;27(2):257-264. doi:

[2.] Cancarevic I, Rehman M, Iskander B, Lalani S, Malik BH. Is There a Correlation Between Irritable Bowel Syndrome and Lactose Intolerance? Cureus. 2020 Jan 20;12(1):e6710. doi: 10.7759/cureus.6710. PMID: 32104635; PMCID: PMC7032600.

[3.] Dukowicz AC, Lacy BE, Levine GM. Small intestinal bacterial overgrowth: a comprehensive review. Gastroenterol Hepatol (N Y). 2007 Feb;3(2):112-22. PMID: 21960820; PMCID: PMC3099351.

[4.] Ghafoor A, Karunaratne T, Rao SSC. Bacterial overgrowth and lactose intolerance: how to best assess. Curr Opin Clin Nutr Metab Care. 2022 Sep 1;25(5):334-340. doi: 10.1097/MCO.0000000000000861. Epub 2022 Jul 16. PMID: 35838278.

[5.] Ghoshal UC. How to interpret hydrogen breath tests. J Neurogastroenterol Motil. 2011 Jul;17(3):312-7. doi: 10.5056/jnm.2011.17.3.312. Epub 2011 Jul 14. PMID: 21860825; PMCID: PMC3155069.

[6.] Losurdo G, Leandro G, Ierardi E, Perri F, Barone M, Principi M, Leo AD. Breath Tests for the Non-invasive Diagnosis of Small Intestinal Bacterial Overgrowth: A Systematic Review With Meta-analysis. J Neurogastroenterol Motil. 2020 Jan 30;26(1):16-28. doi: 10.5056/jnm19113. PMID: 31743632; PMCID: PMC6955189.

[7.] Malik TF, Panuganti KK. Lactose Intolerance. [Updated 2023 Apr 17]. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2024 Jan-. Available from:

[8.] Misselwitz B, Pohl D, Frühauf H, Fried M, Vavricka SR, Fox M. Lactose malabsorption and intolerance: pathogenesis, diagnosis and treatment. United European Gastroenterol J. 2013 Jun;1(3):151-9. doi: 10.1177/2050640613484463. PMID: 24917953; PMCID: PMC4040760.

[9.] Ong DK, Mitchell SB, Barrett JS, Shepherd SJ, Irving PM, Biesiekierski JR, Smith S, Gibson PR, Muir JG. Manipulation of dietary short chain carbohydrates alters the pattern of gas production and genesis of symptoms in irritable bowel syndrome. J Gastroenterol Hepatol. 2010 Aug;25(8):1366-73. doi: 10.1111/j.1440-1746.2010.06370.x. PMID: 20659225.

[10.] Pimentel, Mark MD, FRCP(C), FACG1; Saad, Richard J. MD, FACG2; Long, Millie D. MD, MPH, FACG (GRADE Methodologist)3; Rao, Satish S. C. MD, PhD, FRCP, FACG4. ACG Clinical Guideline: Small Intestinal Bacterial Overgrowth. The American Journal of Gastroenterology 115(2):p 165-178, February 2020. | DOI: 10.14309/ajg.0000000000000501  

[11.] Rana SV, Malik A. Hydrogen breath tests in gastrointestinal diseases. Indian J Clin Biochem. 2014 Oct;29(4):398-405. doi: 10.1007/s12291-014-0426-4. Epub 2014 Mar 21. PMID: 25298621; PMCID: PMC4175689.

[12.] 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. doi: 10.1038/ajg.2017.46. Epub 2017 Mar 21. PMID: 28323273; PMCID: PMC5418558.

[13.] Tahli Singer-Englar, Rezaie A, Gupta K, et al. 182 - Competitive Hydrogen Gas Utilization by Methane- and Hydrogen Sulfide-Producing Microorganisms and Associated Symptoms: Results of a Novel 4-Gas Breath Test Machine. Gastroenterology. 2018;154(6):S-47. doi:

[14.] Tansel A, Levinthal DJ. Understanding Our Tests: Hydrogen-Methane Breath Testing to Diagnose Small Intestinal Bacterial Overgrowth. Clin Transl Gastroenterol. 2023 Apr 1;14(4):e00567. doi: 10.14309/ctg.0000000000000567. PMID: 36744854; PMCID: PMC10132719.

About the Test

The trio-smart breath test measures the levels of hydrogen, methane, and hydrogen sulfide in a patient’s breath to determine if any are elevated abnormally. ​It aids in the identification of small intestinal bacterial overgrowth (SIBO), intestinal methanogenic overgrowth (IMO), and excess hydrogen sulfide. This test comes with a GLUCOSE substrate.

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Lab Test Information
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Lab Company
Gemelli Biotech
Sample Type
Shipping Time
4 - 8 days
Turnaround Time
7 days
Test Preparation Starts
Up to 7 days before collection
Number of Collection Days
1 day
Methods Used For Processing
Gas Chromatography
Lab Certifications
CLIA Certified
CAP Accredited
ISO 15189
COLA Accredited
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