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目的建立快速检测和鉴别单增李斯特菌、绵羊李斯特菌、英诺克李斯特菌、威尔李斯特菌和格氏李斯特菌等5种常见李斯特菌的方法。方法采用PCR-RFLP技术,首先通过引物“Lis1A-Lis1B”对李斯特菌属iap基因进行扩增,扩增产物大小约1.4kb,然后用限制性内切酶DdeⅠ对PCR产物进行酶切,电泳观察具有种间特异性的酶切谱带进行鉴别。结果上述5种李斯特菌的PCR扩增产物经内切酶DdeⅠ消化后,得到片段大小不同、具有种间差异的特异性酶切图谱。结论本实验建立的PCR-RFLP方法可以用于上述5种常见李斯特菌的快速检测和鉴定。
OBJECTIVE To establish a rapid method for the detection and identification of 5 common Listeria monocytogenes, including Listeria monocytogenes, Listeria monocytogenes, Listeria lynx, Listeria monocytogenes and Listeria grisea. Methods PCR-RFLP technique was used to amplify the iap gene of Listeria by primer “Lis1A-Lis1B”. The size of the product was about 1.4kb, then the PCR product was digested with restriction endonuclease DdeⅠ , Electrophoresis with species-specific enzyme-cut band identification. Results The PCR amplification products of the above five species of Listeria were digested with endonuclease Dde Ⅰ to get the specific digestion maps with different sizes and interspecific differences. Conclusion The PCR-RFLP method established in this experiment can be used for the rapid detection and identification of the above five common Listeria.