{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,19]],"date-time":"2026-06-19T05:04:09Z","timestamp":1781845449760,"version":"3.54.5"},"reference-count":18,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2003,9,23]],"date-time":"2003-09-23T00:00:00Z","timestamp":1064275200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Nahrung"],"published-print":{"date-parts":[[2003,10]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Patients with coeliac disease (gluten\u2010sensitive enteropathy) are intolerant against gliadins from wheat and the respective proteins from related cereals and have to keep a lifelong gluten\u2010free diet. For control of gliadin in gluten\u2010free food sensitive assay techniques are necessary. We developed an immunopolymerase chain reaction (iPCR) assay for gliadin. In this technique immunological detection of gliadin by a monoclonal antibody R5 conjugated with an oligonucleotide is amplified by PCR. For quantification, iPCR was performed as real\u2010time PCR (real\u2010time iPCR) in one step. By means of real\u2010time iPCR, the sensitivity of gliadin analysis was increased more than 30\u2010fold above the level reached by enzyme immunoassay. Real time\u2010iPCR using R5 directly conjugated with oligonucleotide was clearly more sensitive than real time\u2010iPCR applying sequentially biotinylated R5, streptavidin, and biotinylated oligonucleotide. With directly conjugated R5 gliadin was detected at a concentration as low as 0.16 ng\/mL corresponding to 16 \u03bcg gliadin\/100 g food or 0.16 ppm (corresponding to 0.25 g of food extracted in 10 mL of solvent and 25\u2010fold dilution of the extract prior to analysis). This is the first report applying the highly sensitive technique of iPCR for gliadin analysis. Furthermore, this is the first approach to perform real\u2010time 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.<\/jats:p>","DOI":"10.1002\/food.200390079","type":"journal-article","created":{"date-parts":[[2003,9,23]],"date-time":"2003-09-23T22:09:34Z","timestamp":1064354974000},"page":"345-348","source":"Crossref","is-referenced-by-count":26,"title":["Assay of gliadin by real\u2010time immunopolymerase chain reaction"],"prefix":"10.1002","volume":"47","author":[{"given":"Nadine","family":"Henterich","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Awad A.","family":"Osman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Enrique","family":"M\u00e9ndez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas","family":"Mothes","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2003,9,23]]},"reference":[{"key":"e_1_2_1_2_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1651-2227.1996.tb14239.x"},{"key":"e_1_2_1_3_2","doi-asserted-by":"publisher","DOI":"10.1097\/00042737-200106000-00023"},{"key":"e_1_2_1_4_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2249.2000.01312.x"},{"key":"e_1_2_1_5_2","doi-asserted-by":"publisher","DOI":"10.1084\/jem.191.4.603"},{"key":"e_1_2_1_6_2","doi-asserted-by":"publisher","DOI":"10.1038\/73200"},{"key":"e_1_2_1_7_2","doi-asserted-by":"publisher","DOI":"10.1053\/gast.2002.33606"},{"key":"e_1_2_1_8_2","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20012028"},{"key":"e_1_2_1_9_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1074129"},{"key":"e_1_2_1_10_2","doi-asserted-by":"publisher","DOI":"10.1097\/01.meg.0000059119.41030.df"},{"key":"e_1_2_1_11_2","doi-asserted-by":"publisher","DOI":"10.1097\/00042737-200110000-00011"},{"key":"e_1_2_1_12_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1439758"},{"key":"e_1_2_1_13_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/21.16.3761"},{"key":"e_1_2_1_14_2","doi-asserted-by":"publisher","DOI":"10.1016\/0009-8981(96)06401-7"},{"key":"e_1_2_1_15_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/22.25.5530"},{"key":"e_1_2_1_16_2","unstructured":"Garc\u00eda E. Llorente M. Hernando A. M\u00e9ndez E. in:Stern M.(Ed.(Proc. 17th Meeting Working Group Prolamin Analysis and Toxicity London Verlag Wissenschaftliche Scripten Zwickau 2003 pp. 35\u201343."},{"key":"e_1_2_1_17_2","first-page":"53","volume":"2","author":"Niemeyer C. M.","year":"1996","journal-title":"Biospektrum"},{"key":"e_1_2_1_18_2","doi-asserted-by":"publisher","DOI":"10.1007\/s002170050306"},{"key":"e_1_2_1_19_2","unstructured":"L\u00f3pez E. Reyes E. Hern\u00e1ez M. Llorente M. M\u00e9ndez E. Wieser H. Proc. 15th Meeting Working Group Prolamin Analysis and Toxicity Meran November 10\u201312 2000 pp. 129\u2013138."}],"container-title":["Food \/ Nahrung"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Ffood.200390079","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/food.200390079","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,11,17]],"date-time":"2023-11-17T00:37:35Z","timestamp":1700181455000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/food.200390079"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2003,9,23]]},"references-count":18,"journal-issue":{"issue":"5","published-print":{"date-parts":[[2003,10]]}},"alternative-id":["10.1002\/food.200390079"],"URL":"https:\/\/doi.org\/10.1002\/food.200390079","archive":["Portico"],"relation":{},"ISSN":["0027-769X","1521-3803"],"issn-type":[{"value":"0027-769X","type":"print"},{"value":"1521-3803","type":"electronic"}],"subject":[],"published":{"date-parts":[[2003,9,23]]}}}