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目的探讨实时荧光PCR(Real-time PCR)技术应用于致病性钩端螺旋体(钩体)快速检测的可行性。方法以钩体rrs基因作为靶基因,合成1对引物和1条Taq Man探针,建立致病性钩体Taq Man Real-time PCR检测方法,进行灵敏性、特异性验证。用建立的方法对90份鼠肾样品进行检测,并与普通PCR方法进行比较。结果建立的致病性钩体Real-time PCR检测方法线性关系较好,灵敏性比普通PCR高100倍,能特异性检出致病性钩体;对鼠肾样品的检测与普通PCR检测结果符合率为97.78%。结论以rrs基因为靶基因建立的Real-time PCR方法时效性好、灵敏度高、特异性强,可用于致病性钩体的快速检测。
Objective To explore the feasibility of Real-time PCR (Real-time PCR) in the rapid detection of pathogenic Leptospira (Leptospira). Methods One pair of primers and one Taq Man probe were synthesized using the rrs gene of Leptospira interrogans. Taq Man Real-time PCR was used to detect the pathogen. Ninety mouse kidney samples were tested by the established method and compared with the normal PCR method. Results The Real-time PCR method established by the real-time PCR method showed a good linearity and the sensitivity was 100 times higher than that of the normal PCR. It could detect pathogenic leptospirosis specifically. The detection of renal samples and ordinary PCR results The coincidence rate is 97.78%. Conclusion The Real-time PCR method using rrs gene as target gene has good timeliness, high sensitivity and specificity. It can be used for the rapid detection of pathogenic leptospira.