16S rRNA荧光定量PCR法检测双歧杆菌

DETECTING BIFIDOBACTERIA BY 16S RRNA FLUORESECENT QUANTIFICATIVE PCR

目的 应用 16 S r RNA序列设计引物定量测定双歧杆菌。方法 应用青春型双歧杆菌 2 6 2 7号作常规 PCR扩增出的模板经系列稀释做荧光定量 PCR,制作标准曲线 ;对青春型、长型、短型、婴儿型、两歧型、链型等 6个种共 15株双歧杆菌和大肠杆菌等 10株其他细菌做荧光定量 PCR,计算机通过与标准曲线比较给出定量结果 ;对青春型双歧杆菌 2 6 2 7号进行敏感性测定。结果 15株双歧杆菌 (10 6 个菌 )荧光定量 PCR法得到拷贝数 (1.0× 10 6 ～ 1.4× 10 6 拷贝 )基本一致。结论 通过荧光定量

Objective:Quantificative detecting Bifidobacteria by 16S rRNA-targeted PCR.Methods:The standard curve of fluorescent quantificative PCR was deveoped by using a series of dilution of PCR products from Bifidobacterium 2627.Compared with the standard curve,15 strains of Bifidobacteria and 10 strains other bacteria were measured by fluorescent quantificative PCR.The sensitivity was detected by fluorescent quantificative PCR of B.adolescentis 2627.Results:The range of fluorescent quantificative PCR of 15 strains of Bifidobacteria(106bacteria)was nearly equal(1.0×106～1.4×106copies).Conclusion:fluorescent quantificative PCR can be applicable for the quantification fo Bifidobadteria.