一种副干酪乳杆菌快速鉴定方法的建立及应用

Establishment and Application of a Rapid Identification Method for Lactobacillus paracasei

目的:基于肠杆菌基因间重复一致序列聚合酶链反应技术(Enterobacterial Repetitive Intergenic Consensus-Polymerase Chain Reaction,ERIC-PCR),寻找并制备一对引物探针,灵敏、准确地从复杂生境中鉴定副干酪乳杆菌。方法:寻找副干酪乳杆菌HP-B1145的肠杆菌基因间重复一致序列(Enterobacterial Repetitive Intergenic Consensus,ERIC)最佳反应体系,对ERIC片段回收纯化得到特定条带及序列结果,据此设计引物,快速鉴定副干酪乳杆菌。结果:根据回收得到的副干酪乳杆菌HP-B1145 ERIC片段,设计出了一对引物探针(1145-S-F:5’-GCAGGATGCTGATTGTTAGCAG-3’;1145-S-R:5’-CTCCGACCAAAGCGACTATGAC-3’)。结论:根据引物特异性、准确性、普适性、含量限度实验得出,设计的引物能准确灵敏地从复杂生境中鉴定副干酪乳杆菌。

Objective:To search for and prepare a pair of primer probes based on ERIC-PCR technology for sensitive and accurate identification of Lactobacillus paracasei from complex habitats.Method:To search for the optimal ERIC reaction system for Lactobacillus paracasei HP-B1145,and obtain specific bands and sequence results through the recovery and purification of ERIC fragments.Based on this,primers were designed to achieve rapid identification of Lactobacillus paracasei.Result:A pair of primer probes(1145-S-F:5’-GCAGGATGCTGATTGTTAGCAG-3’;1145-S-R:5’-CTCCGACCAAAGCGACTATGAC-3’)were designed based on the recovered HP-B1145 ERIC Fragment.Conclusion:Based on the comprehensive experiments of primer specificity,accuracy,universality,and content limit,the designed primers can accurately and sensitively identify Lactobacillus paracasei from complex habitats.