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| Development of PCR-ELISA for identification of Listeria monocytogenes |
| Bing-Qian Tan Qi-gai He Huan-chun Chen |
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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.
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Received: 18 July 2007
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Corresponding Authors:
Bing-Qian Tan
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NI He-Jia, CHENG Xue, WANG Jing, GAO Ze-Ping, WEI Pan-Pan, TAN Jia-Li, SUN Jing, LI Hai-Tao, GAO Ji-Guo. Isolation and Screening of High Virulent Bacillus thuringiensis Strains Against Lepidopteran Pests in Some Areas of Liaoning Province[J]. 农业生物技术学报, 2019, 27(8): 1513-1520. |
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