PCR-RFLP方法检测和鉴别五种李斯特菌

Rapid detection and identification of five Listeria species by PCR-RFLP

中文关键词:  李斯特菌  限制性片段长度多态性聚合酶链反应  检测  鉴定

英文关键词:Listeria species  PCR-RFLP  detection  identification

中文摘要:

      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1.4 kb, 然后用限制性内切酶Dde Ⅰ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Dde Ⅰ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。

英文摘要:

      Objective To establish a method to detect and identify five type strains of Listeria species such as Listeria monocytogenes, L. ivanuii, L. innocua, L. welshimeri and L. grayi. Methods The 1.4 kb gene iap from all members of the genus Listeria was amplified by using specific primers “Lis1A-Lis1B”, then cut by restriction endonuclease DdeⅠ. The different Listeria species could be identified through the specific enzyme spectrum of electrophoresis. Results The PCR amplification product of the five Listeria species were digested by DdeⅠ, and different sizes of fragments and species-specific endonuclease map were gotten. Conclusion This method with PCR-RFLP can be used to detect and identify the five common Listeria species rapidly.