Development of a sandwich ELISA for the detection of Listeria spp. using specific flagella antibodies
2005
Kim, S.H. (National Veterinary Research and Quarantine Service, MAF, Anyang, Republic of Korea) | Park, M.K. (Cheju National University, Jeju, Republic of Korea) | Chuong, Pham Duc (Thainguyen University of Agriculture and Forestry, Thainguyen, Vietnam) | Lee, Y.S. (Seoul National University, Seoul, Republic of Korea) | Yoon, B.S. (Kyonggi University, Suwon, Republic of Korea) | Kim, J.Y. (Cheju National University, Jeju, Republic of Korea) | Hwang, K.K. (Cheju National University, Jeju, Republic of Korea) | Lim, Y.K. (Cheju National University, Jeju, Republic of Korea), E-mail: yklim@cheju.ac.kr
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 E.coli 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 1×10∨5 cells/0.1 ml at 22℃ and 1×10∨6 cells/0.1 ml at 30℃.
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