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目的探讨lipL32-PCR检测方法对钩端螺旋体(钩体)检测的可行性。方法根据外膜脂蛋白基因(lipL32)设计引物,与卫生行业标准推荐的引物G1/G2对比,进行特异性与灵敏度研究,并应用于常山县蛙、鼠肾脏标本钩体PCR检测;对浙江省2008年分离菌株进行钩体PCR鉴定。结果 lipL32-PCR具有较高的灵敏度和特异性,该引物可特异性扩增致病性钩体DNA,对本实验所用的其他细菌不扩增。2008年分离的钩体菌株PCR鉴定结果与血清学鉴定结果相符;鼠和蛙肾标本进行lipL32-PCR和卫生行业标准的G1/G2PCR检测,结果两法符合率为95.0%;lipL32-PCR检测阳性率为10.0%,G1/G2-PCR检测阳性率为5.0%,采用精确计算概率法进行阳性率比较,二者差异无统计学意义(P=0.25)。结论 lipL32-PCR检测方法可灵敏、特异地检测致病性钩体,能正确有效地反映野生动物带菌率,为控制钩体病提供依据。
Objective To investigate the feasibility of using lipL32-PCR to detect Leptospira (Leptospira). Methods The primers of lipL32 gene were designed and compared with the recommended primers G1 / G2 in the health industry to study the specificity and sensitivity. The PCR was applied to the PCR detection of the frog and mouse kidney specimens in Changshan County. 2008 isolates were identified by hook PCR. Results lipL32-PCR with high sensitivity and specificity, the primer can be specifically amplified pathogenic Leptospira DNA, for the other bacteria used in this experiment does not amplify. The PCR results of the isolates of Leptospira isolates from 2008 were consistent with the results of serological identification. The results of lipL32-PCR and G1 / G2 PCR of health industry showed that the coincidence rate of two methods was 95.0% The positive rate of G1 / G2-PCR was 5.0%, and the positive rate was calculated by the exact calculation probability method. The difference was not statistically significant (P = 0.25). Conclusion The lipL32-PCR detection method can sensitively and specifically detect the pathogenic leptospira, which can reflect the carriage rate of wild animals correctly and effectively, and provide the basis for the control of leptospirosis.