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http://dx.doi.org/10.5307/JBE.2012.37.6.420

Development of rapid diagnosis technology for porcine proliferative enteropathy (1) - Preparation of the samples and antibody for rapid detecting the lawsonia in pig feces -  

Kim, Hyuck Joo (National Academy of Agricultural Science, RDA)
Hong, Jong Tae (National Academy of Agricultural Science, RDA)
Yu, Byeong Kee (National Academy of Agricultural Science, RDA)
Kim, Gi Young (National Academy of Agricultural Science, RDA)
Lee, Jin Ju (College of Veterinary medicine, Gyeongsang National University)
Kim, Suk (College of Veterinary medicine, Gyeongsang National University)
Publication Information
Journal of Biosystems Engineering / v.37, no.6, 2012 , pp. 420-428 More about this Journal
Abstract
Purpose: Porcine proliferative enteropathy(PPE), caused by the obligate intracellular bacterium Lawsonia intracellularis, is a widely distributed disease throughout the world causing substantial economic loss. The bacterial pathogen invades the intestinal epithelial cells which causes hyperplasia of the infected cells and leads to the process of disease pathogenesis. For diagnosing PPE in a pig farm in earlier stage, a rapid diagnosing test equipment is needed for farmers. To test the equipment appropriately, we prepare the samples and antibodies for rapid detecting the Lawsonia intracellularis in pig feces. Methods : To prepare the PPE infected samples, we sampled PPE suspected pig feces in a pig farm. To manufacture a anti-Lawsonia intracellularis antibody for capturing the Lawsonia intracellularis, the rabbit-anti LsaA synthetic peptide polyclonal antibody was inoculated to rabbits. To select the couple of antibodies which is most well sandwiched with the bacteria, ELISA test was done with PPE infected ileum samples. Finally, to verify the PPE infected feces which would be used to test the rapid kit, PCR test was done on the sampled PPE suspected feces Results : The rabbit-anti LsaA synthetic peptide polyclonal antibody is developed, and is verified to capture the bacterial well through the fluorescence antibody test. Also, we found that the monoclonal antibody and the polyclonal antibody could be used as couples for sandwiching the bacteria. Finally, through the PCR test for samples of pig feces, we could prepare the 150 PPE positive samples and 50 PPE negative samples. Conclusions : The manufactured polyclonal antibody and the imported monoclonal antibody could be used to capture the bacteria using the sandwich techniques. Also, the prepared PPE infected negative and positive samples could be used to test the performance of the rapid kit to capture the bacterium Lawsonia intracellularis.
Keywords
Porcine proliferative enteropathy (PPE); rapid diagnosis; pig feces; Lawsonia intracellularis;
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Times Cited By KSCI : 2  (Citation Analysis)
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