Abstract：Objective to detect Listeria monocytogenes in foods, a simple PCR method and PCR-ELISA method were developed, according to hlyA gene sequence clone and analysis, specific PCR primers labled with biotin in 5?end of first primer and a specific probe labled with digxin in 3?end were designed by primer 5.0. The results showed that only L. monocytogenes strains were successfully amplified the expected fragment, whereas the E.coli, Salmonella, L.iuanuii, L.innocua, L.innocua, L.seeligeri, and L.gray were negative. Simple PCR method followed by agarose gel electrophoresis and staining of PCR products with ethidium bromide, while PCR-ELISA detected amplified products wih an enzyme-linked oligosorbent assay determinat optical density value at 405 nm. 60 foodstuff samples were further screened to validate the PCR-ELISA procedures by the results compare to the isolation method SN/T0148.1-2005. Positive proportions of molecular methods revealed higher sensitivity than isolation and identification method. It抯 showed that PCR-ELISA improved sensitivity and specificity, and the data revealed the sensitivity of detemination of PCR products 150 folds than agarose gel electrophoresis.
谭炳乾 何启盖 陈焕春. 建立PCR-ELISA方法检测单核细胞增多性李斯特菌[J]. , 2008, 16(4): 0-.
Bing-Qian Tan Qi-gai He Huan-chun Chen. Development of PCR-ELISA for identification of Listeria monocytogenes. , 2008, 16(4): 0-.
