Taqman-MGB探针法检测炭疽杆菌

Rapid Identification of Bacillus anthracis based on Taqman-MGB probles

根据GenBank中炭疽杆菌的主基因组上gyrA基因,毒力因子px01上pagA基因和侵袭因子px02上的CapA基因分别设计引物和对应Taqman-MGB探针,建立了检测炭疽杆菌的实时定量PCR方法。该方法具有良好特异性,仅对炭疽杆菌检测结果为阳性,而与蜡样芽孢杆菌、苏云金杆菌、巨大芽孢杆菌、沙门菌、大肠杆菌、链球菌、铜绿假单孢杆菌无交叉反应,检测灵敏度最低可达100copies/μL。对24份皮毛盲样和120份送检样品检测结果表明建立的定量PCR方法与商业化定量PCR试剂盒的符合率为100%。Taqman-MGB探针方法灵敏、特异、重复性好,可应用于炭疽杆菌的快速诊断与监测。

To develop a real-time fluorenscence quantitative PCR assay with Taqman-MGB proble for rapid identification of Bacillus anthracis. The Taqman MGB probes were designed based on gyrA,pagA and CapA gene,respectively and quantitative PCR were established. The specificity, sensitivity and stability were assessed. The assay was able to accurately detect Bacillus anthracis and no cross reactitvity to Bacillus cereus, Bacillus thuringiensis , Bacillus megaterium , Salmonella, E. coli 0157 ,β- Streptococcus , Pseudomonas aeruginosa. The detection limit was 100 copies/ μL. 24 blind samples and 120 clinical samples were detected by the established assay and commercial quantitative RT-PCR,the results of two methods of was were 100% consistent. The assay is appropriate for the screening of Bacillus anthracis in clinical samples due to its low-cost, high specificity and sensitivity.