【摘要】 由于硫酸盐还原菌(SRB)的生物学多样性以及在系统发育学上的分散性,采用传统方法筛选活性污泥中的SRB准确率不高,且耗时、耗力,而直接采用16S rRNA基因(rDNA)序列分析方法既耗资又不适合从大量细菌中检测SRB.本研究开发出一种新方法,采用SRB16S rDNA特异引物SRB385F为正向引物,真细菌通用引物EUB926R为反向引物,通过梯度PCR,选择适当的退火温度来扩增SRB通用培养基分离的细菌,通过扩增条带与阳性和阴性对照比较,判断待检测菌株是否为SRB,整个过程仅需6h即可完成.将测试菌株进行16S rDNA测序表明该技术获得的阳性菌株全部为SRB.更多还原

【Abstract】 The common way for screening sulfate-reducing bacteria(SRB) from activated sludge is inaccurate,time-and labor-consuming due to the high biodiversity and phylogenetic decentralization of this kind of anaerobe.And 16S ribosomal RNA gene(rDNA) sequencing directly is unsuited to screen SRB out substantive bacteria in view of its cost.In this study,a novel method based on gradient PCR was developed using SRB-specific 16S rDNA primer SRB385F as forward primer and eubacterial universal primer EUB926R as the reverse primer.An improved annealing temperature was found which would amplify the 16S rDNA of the bacteria isolated on SRB universal culture media.Compared the amplified band with positive and negative controls,it can be determined whether the isolates are SRB or not.The procedure requires only about 6 hours.Subsequently,it is demonstrated that all positive strains are SRB via the 16S rDNA sequences determined.更多还原