Tuyển tập các báo cáo nghiên cứu khoa học quốc tế về bệnh thú y đề tài: Development of a sandwich ELISA for the detection of Listeria spp. using specific flagella antibodies | J. Vet. Sci. 2005 6 1 41-46 JOURNAL OF Veterinary Science Development of a sandwich ELISA for the detection of Listeria spp. using specific flagella antibodies Seong-Hee Kim1 Min-Keun Park2 Jin-Young Kim2 Pham Duc Chuong3 Yong-Soon Lee4 Byoung-Su Yoon5 Kyu-Kye Hwang2 Yoon- Kyu Lim2 National Veterinary Research and Quarantine Service Ministry of Agriculture and Forestry Anyang 430-824 Korea Department of Veterinary Medicine Cheju National University Jeju 690-756 Korea Faculty of Animal Science and Veterinary Medicine Thainguyen University of Agriculture and Forestry Thainguyen Vietnam Department of Veterinary Medicine Seoul National University Seoul 151-742 Korea Department of Biology College of Natural Science Kyonggi University Suwon 442-760 Korea Five monoclonal antibodies MAbs and chicken immunoglobulin IgY were developed by immunizing with flagella purified from Listeria monocytogenes 4b and the five MAbs have been confirmed to be specific against three different epitopes of flagellin. The antibodies showed specific reaction to Listeria genus and no cross-reactivity with other bacteria tested in this experiment including O157 H7 and Salmonella enteritidis. Sandwich enzyme-linked immunosorbent assays ELISA using the MAbs and IgY were developed to detect Listeria species and the sensitivity and specificity of the developed ELISA have been analyzed. The detection limit of ELISA using MAb 2B1 and HrP labeled IgY was 1X 105 cells ml at 22oC and 1X 106 cells ml at 30oC. ELISA using the pair of MAbs MAbs 2B1 and HRP labeled MAbs 7A3 detected up to 104 cells ml at 22oC and 30oC. Detection limit of sandwich ELISA using IgY was 10 times lower than MAb pair. Using the developed ELISA we could detect several Listeria contaminated in food samples after 48 h-culturing. In conclusion both MAbs and IgY have been proved to be highly specific to detect Listeria flagella and the developed sandwich ELISA using these antibodies would be useful tool for screening
