基于FemAB基因特异引物的动物双歧杆菌PCR检测方法的建立

Development of A PCR method for detection of Bifidobacterium animalis based on FemAB gene

分离和鉴定了两株双歧杆菌,通过16SrDNA通用引物扩增条带测序和非特异引物随机扩增法扩增条带测序推测其为动物双歧杆菌。随后使用以编码肽聚糖肽桥形成蛋白(Peptidoglycan bridge formation protein)FemAB基因为目的扩增片段设计的特异引物进行了扩增鉴定。结果表明,此特异引物对这两株动物双歧杆菌菌株扩增结果为阳性,其他对照菌株扩增结果为阴性,且扩增条带测序结果与预计一致。FemAB基因在PCR特异扩增法鉴定细菌种属中具有较好的分辨能力,并使用其建立了动物双歧杆菌PCR检测方法。本研究旨为应用FamAB基因鉴定细菌种属以及双歧杆菌属微生物的鉴定提供了数据参考。

In this experiment, two strains of Bifidobacterium animalis were isolated and speculated by 16S rDNA universal primer amplification and non-specific primer random amplification. Two Bifidobacterium animalis strains were identified finally by the specific primers designed for the peptidoglycan bridge formation protein FemAB gene. The results showed that the specific primers were positive for the two strains of B/fi- dobacterium animalis strains, the results of the other control strains were negative, and the results were consistent with the expected results. The results showed that FemAB gene had enough ability to distinguish bacteria in PCR-specific method, and the PCR detection method of B/fi- dobacterium animalis was established. This study provides a reference for the identification of bacterial species and other Bifidobacterium species by the FamAB gene specific primers.