Fecal Zonulin

Fecal Zonulin

Zonulin has been used as a biomarker of impaired gut barrier function in autoimmune, neurodegenerative, and metabolic diseases

Who Should Be Testing Fecal Zonulin levels?

Any person with autoimmune disease or immune-mediated conditions like:

Autoimmune diseases like:

Fatigue, Low Energy

Celiac disease

Leaky Gut

Ankylosing spondylitis

Inflammatory bowel disease (IBS, IBD, SIBO, Chron’s Colitis)

Type 1 diabetes

Rheumatoid arthritis

Systemic lupus erythematous

Neurological diseases

Multiple sclerosis

Chronic inflammatory demyelinating polyneuropathy




Brain (gliomas)


Lung adenocarcinoma

Ovarian, PCOS




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This stool panel is a comprehensive collection of microbial targets as well as immune and digestive markers. It screens for pathogenic bacteria, commensal bacteria, opportunistic pathogens, fungi, viruses, and parasites. It primarily uses multiplex, automated, DNA analysis to give integrative and functional medicine practitioners a better view into the gastrointestinal microbiome.

Zonulin is a protein secreted by intestinal cells that regulates intercellular tight junctions (1, 2). Tight junctions are the connections between epithelial cells that make up the gastrointestinal lining. Zonulin increases intestinal permeability in the jejunum and ileum (3) and is considered a biomarker for barrier permeability (1, 2). Tight junctions can be opened or closed, depending on the physiological need. Zonulin’s role is to open tight junctions in the gut. In the case of enteric infections, high zonulin can “open the floodgates” and flush out bacteria and toxins (1). Certain gut bacteria and gliadin (the main staple protein from wheat) can activate the zonulin system (2, 4).

The intestinal barrier is a critical interface between the lumen of the gut and the internal milieu. Dysfunction of this barrier is believed to initiate immune dysfunction because it allows macromolecules from the gut lumen to pass into the bloodstream (5). Intestinal permeability, also known as “leaky gut,” has been associated with inflammatory bowel disease, celiac disease, food allergy, irritable bowel syndrome, critical illness, autoimmune diseases (6) and obesity and metabolic disease (7). In many cases, permeability precedes disease (1).

Zonulin regulates barrier permeability. Serum zonulin correlates with intestinal permeability and lactulose/mannitol tests for intestinal permeability (3, 8). High serum zonulin has been associated with celiac disease, type 1 diabetes (8) insulin resistance and type I diabetes (3), cancers, neurological conditions, and autoimmune diseases (see Table 1) (1).


1. Fasano A. Intestinal permeability and it’s regulation by zonulin: diagnostic and therapeutic implications. Clinical gastroenterology and hepatology: the official clinical practice journal of the American Gastroenterological Association. 2012;10(10):1096-1100.

2. Lamprecht M, Bogner S, Schippinger G, et al. Probiotic supplementation affects markers of the intestinal barrier, oxidation, and inflammation in trained men; a randomized, double-blinded, placebo-controlled trial. Journal of the International Society of Sports Nutrition. 2012;9(1):45.

3. Stenman LK, Lehtinen MJ, Meland N, et al. Probiotic With or Without Fiber Controls Body Fat Mass, Associated With Serum Zonulin, in Overweight and Obese Adults-Randomized Controlled Trial. EBioMedicine. 2016;13:190-200.

4. Fasano A, Sapone A, Zevallos V, Schuppan D. Nonceliac gluten sensitivity. Gastroenterology. 2015;148(6):1195-1204.

5. Fasano A. Leaky gut and autoimmune diseases. Clinical reviews in allergy & immunology. 2012;42(1):71-78.

6. Fasano A. Physiological, pathological, and therapeutic implications of zonulin-mediated intestinal barrier modulation: living life on the edge of the wall. The American journal of pathology.2008;173(5):1243-1252.

7. Bischoff SC, Barbara G, Buurman W, et al. Intestinal permeability–a new target for disease prevention and therapy. BMC gastroenterology. 2014;14:189.

8. Wang L, Llorente C, Hartmann P, Yang AM, Chen P, Schnabl B. Methods to determine intestinal permeability and bacterial translocation during liver disease. J Immunol Methods.2015;421:44-53.

9. Wang W, Uzzau S, Goldblum SE, Fasano A. Human zonulin, a potential modulator of intestinal tight junctions. Journal of cell science. 2000;113 Pt 24:4435-4440.

10. Lamprecht M, Bogner S, Steinbauer K, et al. Effects of zeolite supplementation on parameters of intestinal barrier integrity, inflammation, redoxbiology and performance in aerobically trained subjects. Journal of the International Society of Sports Nutrition. 2015;12:40.

Collection Instructions

Step 1 – Please write your name and date of birth on the capped vial.

Step 2 – If possible void urine prior to collecting stool. Collect stool by passing stool onto Collection tray.

Step 3 – Using the spoon attached to the cap of the vial, spoon stool from different areas of the sample into the vial. Fill vial to the red fill line. Just over half full.

Step 4 – Carefully mix stool and fluid with the spoon. Replace cap tightly and shake the vial vigorously for 30 seconds.

Step 5 – Place cap vial into ziplock specimen bag along with absorbent pad. Seal the bag. Place the specimen bag with the sample vial into the kit box.

Step 6 – Place Test Request Form into the box with the sample and ship to the laboratory. See shipping instructions below. **

**If you cannot ship the specimen on the day of the collection please refrigerate the sample by placing the box containing the sample into the refrigerator.

Shipping Instructions

Specimens may be shipped Monday through Friday. The lab receives specimens 5 days a week. We only require that the specimen be received within 6 days after collection.

Before shipping is sure that the capped vial and the Test Request Form are labeled and completely filled out. Be sure the sample vial sealed in the ziplock bag and that the Test Request Form is in the Kit box.

Locate the FedEx Clinical Pak mailer. Fill in your name and address on the shipping label attached to the outside of the mailer.

Place kit box into FedEx Clinical Pak. Remove strip to reveal sticky film and press both sides of mailer together to seal the pouch.

Call FedEx to schedule a pick-up. Dial 1-800-238-5355. When the automated greeting begins say “Rep”. When asked if you are shipping a package say “Yes”. A live person will then answer to help schedule your pickup. Let them know you are shipping using a Billable Stamp.


Can you ship your specimen on Friday?

Yes, it is ok to ship specimens to the lab on Friday. The lab receives specimens 6 days a week. We only require that the specimen be received within 6 days after collection.


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