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J Autoimmun. 2004 Feb;22(1):65-72 Celiac.com 01/29/2004 - A new cloning technique developed by Italian researchers may lead to more accurate diagnoses of celiac disease in borderline patients, including those who are asymptomatic. The technique screens for anti-tTG antibodies in the intestinal mucosa by utilizing a cloning process to amplify the antibodies, thus allowing for their detection even in cases where only minute amounts are present. The new technique is similar to that developed and long utilized by Dr. Kenneth Fine of Enterolab, in that both techniques look for the presence of antibodies in the intestinal mucosa rather than in the blood. The new technique also has the potential to easily screen large numbers of people, which, if the researchers are correct, will lead to a celiac disease diagnostic explosion, as those who are missed by current screening methods will be properly diagnosed. The number of celiacs who are missed using current screening techniques is a topic of debate, and Dr. Fines methods have demonstrated that "in normal people without specific symptoms or syndromes , the stool test is just under three times more likely to be positive than blood tests," as reported in the Winter 2004 edition of Scott-Free newsletter. It would be very interesting to see how many people test positive in a healthy population using this new technique. Below is the abstract of the article: One-step cloning of anti tissue transglutaminase scFv from subjects with celiac disease . Celiac disease is characterized by intestinal mucosal injury and malabsorption precipitated by dietary exposure to gluten of some cereals with a prominent role being played by gliadins, specific antigenic determinants found in wheat gluten. Patients suffering from celiac disease have serum antibodies recognizing gliadin, as well as the Endomysial autoantigen tissue transglutaminase. Phage display antibody libraries have revealed ectopic production of anti-transglutaminase antibodies by intestinal lymphocytes with a biased use of the VH5 antibody gene family. Here we report a study on the pairing of VH and VL families in the antibodies to transglutaminase. Our results led to the construction of small phage display antibody libraries based on the amplification of the two genes in the VH5 family from intestinal lymphocytes. This method can be used for the rapid characterization of the anti-transglutaminase response in a potentially large number of subjects including asymptomatic patients whose serum antibodies may be undetectable.
Nahrung. 2003 Oct;47(5):345-8. Celiac.com 01/14/04 – German researchers have developed a new test to determine the level of gliadin, the portion of gluten that is toxic to celiac patients, in foods. This new technique is called immunopolymerase chain reaction (iPCR), and it utilizes immunological detection of gliadin by a monoclonal antibody R5 conjugated when an oligonucleotide is amplified by PCR. The technique yields a "30-fold above the level reached by enzyme immunoassay" in laboratory tests, and it detects concentrations in food "as low as 0.16 ng/ml corresponding to 16 microgram gliadin/100 g food or 0.16 ppm (corresponding to 0.25 g of food extracted in 10 ml of solvent and 25-fold dilution of the extract prior to analysis)." This is the first time that this highly sensitive technique has been used for gliadin analysis, and "is the first approach to perform real-time iPCR in one step without changing the reaction vessels after enzyme immunoassay for subsequent PCR analysis thus minimizing risks of contamination and loss of sensitivity."