Jump to content
This site uses cookies. Continued use is acceptance of our Terms of Use and Privacy Policy. More Info... ×
  • Welcome to Celiac.com!

    You have found your celiac tribe! Join us and ask questions in our forum, share your story, and connect with others.




  • Celiac.com Sponsor (A1):



    Celiac.com Sponsor (A1-M):


  • Get Celiac.com Updates:
    Support Our Content
    eNewsletter
    Donate
  • Scott Adams
    Scott Adams

    Interpretation of Celiac Disease Blood Test Results

    Reviewed and edited by a celiac disease expert.
    Interpretation of Celiac Disease Blood Test Results - Images: CC--Scott Robinson
    Caption: Images: CC--Scott Robinson

    The following detailed explanation of serological tests for celiac disease was written by Tom Ryan, Technical Service Specialist, INOVA Diagnostics, Inc.

    There has been a lot of discussion about serological testing for celiac disease recently, specifically regarding tTG (tissue Transglutaminase) testing. I will try to answer some of the many questions that have appeared on this list about all of the tests. First, and this applies to any of the blood tests, you must currently be on a gluten containing diet for the tests to be accurate. Antibodies are produced by the immune system in response to substances that the body perceives as threatening. The immune response that your body produces is its response to being exposed to gluten in the diet and its subsequent effect on the intestinal mucosa. If there is no gluten in the diet, then there is no response that we can measure. A brief change in diet will not have a noticeable effect. If you have been gluten free for a week or so, it will not make any great difference. The response might be marginally less but the difference is insignificant because the body has not had time to respond to the change. Conversely, if you have been gluten free for a protracted period of time and decide to be tested, a brief challenge of a couple of weeks is not enough to elicit a response and get an accurate test.

    Celiac.com Sponsor (A12):
    There are several steps that take place to generate an immune response and it takes time both for the positive reaction when gluten is present and to clear the antibodies when gluten is eliminated. There has been a great deal of discussion about how much and how long a challenge should be and there is no consensus. Talk with your Doctor. My personal feeling is that the minimum is 2 slices of bread per day for 6 weeks to get an accurate test but I would not try to second-guess the Doctor. There are basically four tests that can be performed to aid in diagnosing celiac disease. Notice that I say they will aid in diagnosing celiac disease. Immunology is fairly accurate but it is far from being an exact science. All of the lab tests, regardless of the type or source, are presented as aids to diagnosis. They should not be used alone as a basis for diagnosis but rather are intended to be considered in conjunction with the physical examination of the patient as well as the reported symptoms, etc. by a trained physician. There has been a great deal of confusion about what the tests are and I hope to alleviate some of the misunderstandings. There are many terms that we hear. tTG, IgA, IgG, ELISA, etc. What are all of these? Some contributors to the list make reference to the IgA or IgG test or to the ELISA test. These labels are incomplete for our purposes and could be referring to any number of different tests.

    We all have, within our bodies, a family of closely related although not identical proteins which are capable of acting as antibodies. These are collectively referred to as immunoglobulins. Five major types of immunoglobulins are normally present in the human adult. They are IgG, IgA, IgM, IgE and IgD. Each of these is a shorthand way of writing immunoglobulin gamma G (or A or M, etc.) and they each perform a different function in our systems. IgG is the principal immunoglobulin in human serum. It is important in providing immunity in a developing fetus because it will pass across the placental barrier. IgA is the principal immunoglobulin in secretions from respiratory and intestinal mucosa. IgE is a gamma globulin produced by cells lining the intestinal and respiratory tracts. It produces the antibodies associated with most hypersensitivity (allergic) responses. It is associated with asthma, hay fever, etc. IgM is a globulin formed in almost every immune response in the early part of the reaction. IgD is a rare protein present in normal sera in a tiny amount. These designations refer to the type of protein that is carrying the antibody in question. Both IgG and IgA subtypes of anti-gliadin antibody are produced, hence we refer to them as IgG gliadin or IgA gliadin. Collectively they are anti-gliadin antibodies.

    Anti-Gliadin Antibodies:

    Both IgA and IgG anti-gliadin antibodies (AGA) are detected in sera of patients with gluten sensitive enteropathy (celiac disease). IgG anti-gliadin antibodies are more sensitive but are less specific markers for disease compared with IgA class antibodies. IgA anti-gliadin antibodies are less sensitive but are more specific. In clinical trials, the IgA antibodies have a specificity of 97% but the sensitivity is only 71%. That means that, if a patient is IgA positive, there is a 97% probability that they have celiac disease. Conversely, if the patient is IgA negative, there is only a 71% probability that the patient is truly negative for celiac disease. Therefore, a positive result is a strong indication that the patient has the disease but a negative result does not necessarily mean that they don not have it. False positive results are rather uncommon but false negative results can occur. On the other hand, the IgG anti-gliadin antibodies are 91% specific and have an 87% sensitivity. This means that they will show positive results more readily but there is not as strong a correlation with celiac disease. It is less specific. Patients with other conditions but not afflicted with celiac disease will occasionally show positive results. IgG anti-gliadin antibodies are detectable in approximately 21% of patients with other gastrointestinal disorders. This test might yield false positive results but is less likely to yield false negative results.

    A sensitive testing protocol includes testing for both IgA and IgG anti-gliadin antibodies since a significant portion of celiac patients (approx. 2-5%) are IgA deficient. This combined IgA and IgG anti-gliadin antibody assay has an overall sensitivity of 95% with a specificity of 90%. The type of test used to detect the anti-gliadin antibodies is called an ELISA. This is an acronym and it stands for Enzyme Linked Immuno-Sorbent Assay. ELISA is not a test in itself. It is a method of testing and it is a relatively simple test to perform. It involves putting a measured amount of diluted patient serum into the wells of a specially constructed and prepared plate and incubating it for a period of time with various chemicals. The end result is a color change, the intensity of which is dependent upon the concentration of anti-gliadin antibody (or other protein being measured) in the patient serum. The ability of this colored solution to absorb light at a particular wavelength can be measured on a laboratory instrument and mathematically compared with solutions that contain a known amount of anti-gliadin antibody to arrive at a number for the amount of antibody present. The sample can then be classified as negative, (0-20 units); weak positive, (21-30 units); or moderate to strong positive if greater than 30 units. The purpose of testing for anti-gliadin antibodies includes, in addition to diagnosis of gluten sensitive enteropathy, monitoring for compliance to a gluten free diet. IgA gliadin antibodies increase rapidly in response to gluten in the diet and decrease rapidly when gluten is absent from the diet. The IgA anti-gliadin antibodies can totally disappear in 2-6 months on a gluten free diet, so they are useful as a diet control. By contrast, IgG anti-gliadin antibodies need a long time, sometimes more than a year, to become negative. The reverse is also true. That is, a patient with celiac disease who has been on a gluten free diet and tests negative for IgA anti-gliadin antibodies, will show a rapid increase in antibody production when challenged by gluten in the diet. Approximately 90% of challenged patients will yield a positive IgA anti-gliadin result within 14-35 days after being challenged. The IgG antibodies are somewhat slower.

    Endomysial Antibodies:

    IgA class anti-endomysial antibodies (AEA) are very specific, occurring only in celiac disease and DH. These antibodies are found in approximately 80% of patients with DH and in essentially 100% of patients with active celiac disease. IgA endomysial antibodies are more sensitive and specific than gliadin antibodies for diagnosis of celiac disease. Antibody titers (dilutions) are found to parallel morphological changes in the jejunum and can also be used to reflect compliance with gluten-free diets. Titers decrease or become negative in patients on gluten free diets and reappear upon gluten challenge.

    The test for anti-endomysial antibodies is more subjective and more complicated for the lab to perform than the anti-gliadin assays. It involves serially diluting some of the patients serum, that is, diluting it by ½ then ¼, 1/8, 1/16, etc. and putting these dilutions on a glass slide that has some sort of tissue affixed to it. The slide is then processed with various solutions and examined under a fluorescent microscope to determine if any of that serum binds to any of the proteins in the tissue. If so, then that patient is confirmed as having antibodies to that particular protein. This method of testing is called an IFA or sometimes IIFA. It stands for Indirect Immuno-Fluorescent Assay. The selection of which tissue slide to use is determined by what specific protein, hence which antibody, you are specifically looking for. Endomysial antibodies react with the endomysium, which is a sheath of reticular fibrils that surround each muscle fiber. Therefore, to detect endomysial antibodies, you would want to use a tissue substrate that contains a lot of muscle tissue. The substrate used most often for this assay is distal sections of the esophagus. These are very thinly sliced and fixed to the slide. They contain muscle fibers and not much else so there is a lot of endomysium available to react with the anti-endomysial antibodies.

    Reading this test involves viewing the reacted slides with a fluorescent microscope to make the determination. This requires a highly skilled and trained eye and, of necessity, is somewhat subjective. You are looking for a green fluorescence in the endomysium covering the muscle fibers. The test is reported as the titer or final dilution in which the fluorescence can still clearly be seen. As you can imagine, this is very subjective. There are no standardized values and it is up to the judgment of the particular technician what the endpoint titer is. Recently, (1998) the endomysial antigen targeted by the anti-endomysial antibodies was identified as the protein cross-linking enzyme known as tissue transglutaminase (tTG). This has enabled the production of an antigen specific ELISA assay incorporating tTG as a reliable and objective alternative to the traditional and subjective Immunofluorescence based assays. In clinical trials, the correlation with the endomysial IFA assay has been shown to be close to 100%. This is a test that has been very well received in the professional community. It is an ELISA, like the anti-gliadin antibody test and, as such, is not subject to interpretation like the IFA. That is the greatest advantage to this new test! With this or any ELISA, the response is measured on an instrument that calculates the amount of light of a particular wavelength that is absorbed by the solution and prints out a numerical result. There is no chance of human error skewing the results because there is no judgment call involved. The ELISA plate, regardless of what you are testing for, is processed with at least three control sera (sometimes as many as eight) in addition to the unknown sample being tested. There is a negative serum and at least two positive sera containing different levels of the antibody being tested. There are specific requirements for the absorption levels of these three controls. That is, each of them has a minimum or maximum (or both) number that must be seen by the instrument in order for it to be a valid test. If there is any variance from these expected numbers, it is an indication that something went wrong and the test results are discarded and the test repeated. There is therefore no way the technician could report inaccurate results, (assuming they diluted the sample correctly). Either the test was valid, and you can rely upon the accuracy of the result, or the test is invalid, and the entire result discarded. If any error was made during the processing of the ELISA plate, it would result in the control sera numbers being out of range and the entire test result would be thrown out.

    In summary, the tTG ELISA is measuring the same thing that the endomysial IFA is measuring but with a method that is more sensitive and specific and not subject to interpretation. IgA class Reticulin antibodies are found only in Celiac disease and dermatitis herpetiformis. These antibodies are found in approximately 60% of celiac disease patients and 25% of DH patients. This test is falling into disuse because of the limited utility and the availability of better tests. It is an IFA performed on a tissue substrate with all the attendant problems that go along with it. The development of all of these serum assays has tremendously simplified the diagnosis of celiac disease and improved the accuracy as well. The original criteria for diagnosis according to the European Society for Pediatric Gastroenterology and Nutrition, (ESPGAN), involved a year of arduous studies with:

    • An initial positive gut biopsy;
    • 6 months on a gluten free diet;
    • A second, negative gut biopsy;
    • A gluten challenge for 6 months and;
    • A third, positive gut biopsy. The revised ESPGAN criteria call for positive results in two of the serological tests confirmed by a single positive biopsy. In practice, many gastroenterologists are utilizing the serologies in conjunction with a controlled diet and the clinical presentation to form a basis for diagnosis without the need for the invasive procedure.

    Through the auspices of the Celiac Disease Foundation and others, a professional symposium and workshop was organized earlier this year in Marina Del Rey, California with participants from Europe as well as the U.S. to establish standards for reporting test results. This should improve testing and diagnosis even more. At the conclusion of this conference a Celiac Disease Standardization Committee was formed to investigate and make recommendations on a standardized method of reporting results.



    User Feedback

    Recommended Comments



    Guest Phillipa Sheard

    Posted

    In order for you to get a true test result you can not eat out. Gluten is in everything. Dry beans are persevered with Wheat, as Scott Adams said to me a couple of days " The Food May Be Cross Contaminated" Try to eat veggies,or anything you feel in manufactures can be cross contaminated with Gluten and then go take another Panel.

    Link to comment
    Share on other sites
    Guest admin
    In order for you to get a true test result you can not eat out. Gluten is in everything. Dry beans are persevered with Wheat, as Scott Adams said to me a couple of days " The Food May Be Cross Contaminated" Try to eat veggies,or anything you feel in manufactures can be cross contaminated with Gluten and then go take another Panel.

    I don't agree that wheat is used to preserve dry beans, and do believe that you can eat out safely if you take basic steps to discuss your meal with the wait staff and cooks.

    Scott

    Link to comment
    Share on other sites
    Guest Chika
    Finally an explanation! I was low on my IgA but high positive on IgG and so sick and my doc said, 'at some point in the past, gluten made you very sick, but you're fine now.'.. He was totally ignoring my symptoms! That was two years ago and he still doesn't believe I have it, even though since then he diagnosed me with osteopenia.. I've been off gluten for about 3 months now, and my chronic migraines and stomach problems are gone! my doc still doesn't believe I have it, but IBS instead! give me a break! And these are Seattle gastroenterologists!

    Same situation with me and my docs in NJ!

    Link to comment
    Share on other sites
    Guest Debra

    This helped me somewhat. Only have iga of 5.8 and negative on igg and everything else?

    Link to comment
    Share on other sites
    Guest ellen

    Helped somewhat.

     

    Can someone help me interpret 10 year old sons results though? Gliadin AB IGG 5 (says negative), IGA 18 (says >17 positive). Is this automatic Celiac?

    Link to comment
    Share on other sites
    Guest d. finlayson MD

    Posted

    Good job on your article.

    There are some articles on TTG that you might want to look at.

    I could email them to you.

    Link to comment
    Share on other sites
    Guest amii

    I have been diagnosed with celiac disease-I am 14 years old. Celiac doesn't run in my family, but thyroid cancer runs on my dads side, is this connected?

    Link to comment
    Share on other sites
    Guest PeggyNC

    Posted

    terrific article...wish I had known about it before the doctor ordered the blood tests....been gluten free for 5 years after diagnosis in another hospital in western part of the state, so of course they came up negative when current doctor in new location tested. Will print a copy to take with me to the doctor's at next appointment..wish me luck!

    Link to comment
    Share on other sites
    Guest michele

    Posted

    My sons are both coming back with elevated IGA tests the older 165.0 TTG IGA 16 but his symptoms are mood, weight, stuff like that. My younger son can't keep food down and is nauseated all the time his is only 117 and 10. He is miserable they are telling him not to change his diet it might mess up testing but he has been very ill for 3 months and we are frustrated with all the waiting around.

    Any advice?

    Link to comment
    Share on other sites
    Guest Cathy Simms

    Posted

    I was hoping to find an explanation for IGA, Qn. This test is part of the celiac panel and I have not been able to find what an elevated result means.

    Other than the exclusion of this important IGA explanation, the article was very good.

    Link to comment
    Share on other sites
    Guest Jo

    Very good article for us laymen. I am having the testing done in 2 days. I wasn't told whether or not to fast for the test.

    Link to comment
    Share on other sites
    Guest Rosalie Southern

    Posted

    Very informative just had a blood test and I cant really interpret results: antibody part waiting on part 2 Genetic one.

    Link to comment
    Share on other sites



    Create an account or sign in to comment

    You need to be a member in order to leave a comment

    Create an account

    Sign up for a new account in our community. It's easy!

    Register a new account

    Sign in

    Already have an account? Sign in here.

    Sign In Now

  • Get Celiac.com Updates:
    Support Celiac.com:
    Donate
  • About Me

    Scott Adams

    Scott Adams was diagnosed with celiac disease in 1994, and, due to the nearly total lack of information available at that time, was forced to become an expert on the disease in order to recover. In 1995 he launched the site that later became Celiac.com to help as many people as possible with celiac disease get diagnosed so they can begin to live happy, healthy gluten-free lives.  He is co-author of the book Cereal Killers, and founder and publisher of the (formerly paper) newsletter Journal of Gluten Sensitivity. In 1998 he founded The Gluten-Free Mall which he sold in 2014. Celiac.com does not sell any products, and is 100% advertiser supported.


  • Celiac.com Sponsor (A17):
    Celiac.com Sponsor (A17):





    Celiac.com Sponsors (A17-M):




  • Related Articles

    Scott Adams
    Great Smokies Diagnostic Laboratory (GSDL), a private, rapid-growth Functional Medicine Clinical laboratory, announced today receipt of 510(K) market clearance from the Food and Drug Administration (FDA) for its Intestinal Permeability test kit, utilizing the lactulose-mannitol challenge drink. Used in the non-invasive assessment of intestinal permeability, the test demonstrated its superior sensitivity as compared to the existing d-xylose test in measuring intestinal permeability, a measurement used in the diagnosis of gastrointestinal malabsorption syndromes, such as celiac disease, colitis, Crohns disease, and Irritable Bowel Syndrome.
    What is intestinal permeability?
    Intestinal permeability refers to impairment of the intestinal mucosal barrier, which is central to...


    Dr. Rodney Ford M.D.
    This question, “how early can you diagnose celiac disease?” is a major concern for both parents and paediatricians.  This is because, like many diseases, celiac disease comes on slowly.  This means that it can take a long time to make the diagnosis.
    Celiac disease can develop slowly?
    Yes, celiac disease can develop very slowly.  The symptoms can be subtle.  It is a progressive disease.  When you are first born, you cannot have celiac disease as you have never been exposed to gluten.  However, if you have the right genetic make up (that is you have the celiac gene) and the right environmental circumstances (eating gluten and getting gut inflammation), then celiac disease can develop.
    Finding tissue damage
    Celiac disease is a condition that is recognised when you ...


    Jefferson Adams
    Celiac.com 08/11/2009 - While the use of anti-tTG antibodies is common practice in the diagnosis of celiac disease, their value in long-term follow-up remains controversial. A team of researchers recently set out to assess the value of anti-tTG antibodies in long-term follow-up.
    The research team was made up of C.R. Dipper, S. Maitra, R. Thomas, C.A. Lamb, A.P.C. McLean-Tooke, R. Ward, D. Smith, G. Spickett, and J.C. Mansfield. Their goal was to see if they could use serial anti-tTG antibody levels to gauge adherence to a gluten-free diet (GFD) and to spot patients facing complications from celiac disease.
    Researchers conducted a cohort follow-up study of 182 adult subjects over 54-months. The team charted patient self-assessment of gluten-free diet adherence; anti-tTG antibody concentration...


  • Recent Activity

    1. - Scott Adams replied to Jack Common's topic in Celiac Disease Pre-Diagnosis, Testing & Symptoms
      3

      How many grams is there in one slice of wheat bread?

    2. - Jack Common replied to Jack Common's topic in Celiac Disease Pre-Diagnosis, Testing & Symptoms
      3

      How many grams is there in one slice of wheat bread?

    3. - Scott Adams replied to Barcino's topic in Post Diagnosis, Recovery & Treatment of Celiac Disease
      2

      Antibodies went up

    4. - Scott Adams replied to Jack Common's topic in Celiac Disease Pre-Diagnosis, Testing & Symptoms
      3

      How many grams is there in one slice of wheat bread?

    5. - Scott Adams replied to Kathleen JJ's topic in Traveling with Celiac Disease
      1

      Ski trip with Celiac son - preparation advice please


  • Celiac.com Sponsor (A19):



  • Member Statistics

    • Total Members
      126,378
    • Most Online (within 30 mins)
      7,748

    Kelvin Chola
    Newest Member
    Kelvin Chola
    Joined

  • Celiac.com Sponsor (A20):


  • Forum Statistics

    • Total Topics
      120.9k
    • Total Posts
      69.4k

  • Celiac.com Sponsor (A22):





  • Celiac.com Sponsor (A21):



  • Popular Now

    • disneyfamilyfive
      4
    • BoiseNic
      10
    • Jtestani
  • Popular Articles

    • Scott Adams
    • Scott Adams
    • Scott Adams
    • Scott Adams
    • Scott Adams
  • Upcoming Events

×
×
  • Create New...