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目的建立鉴定乳酸杆菌及嗜热链球菌的16SrRNA基因PCR方法。方法优化细菌DNA提取方法及PCR反应条件,将细菌16SrRNA基因的PCR产物克隆到质粒载体上并进行序列分析,以对乳酸杆菌酸奶分离株进行鉴定。结果 16SrRNAPCR鉴定方法将50株益生菌酸奶分离株分为7个种,分别为:保加利亚乳酸杆菌24株、嗜热链球菌12株、嗜酸乳酸杆菌7株、干酪乳酸杆菌3株、德氏乳酸杆菌2株、发酵乳酸杆菌1株及巴黎链球菌1株。结论建立的16SrRNA基因PCR方法灵敏、可靠,适用于乳酸杆菌及嗜热链球菌的鉴定。
Objective To establish a 16S rRNA gene PCR method for identification of Lactobacillus and Streptococcus thermophilus. Methods The bacterial DNA extraction method and PCR reaction conditions were optimized. The PCR product of bacterial 16S rRNA gene was cloned into the plasmid vector and sequenced to identify the Lactobacillus yogurt isolates. Results Sixteen strains of probiotic yoghurt were divided into seven species, including 24 strains of Lactobacillus bulgaricus, 12 strains of Streptococcus thermophilus, 7 strains of Lactobacillus acidophilus, 3 strains of Lactobacillus casei, 2 strains of bacillus, 1 strain of Lactobacillus fermentum and 1 strain of Streptococcus aureus. Conclusion The established 16S rRNA gene PCR method is sensitive and reliable and is suitable for the identification of Lactobacillus and Streptococcus thermophilus.