lipL32-PCR应用于钩端螺旋体检测  

Application of a lipL32-PCR method for detection of Leptospira interrogans

作　　者：姜理平[1] 陆群英[1] 罗芸[1] 叶菊莲[1] 张政[1] 徐宝祥[1] 

机构地区：[1]浙江省疾病预防控制中心微生物所,杭州310051

出　　处：《中国媒介生物学及控制杂志》2011年第4期388-391,共4页Chinese Journal of Vector Biology and Control

摘　　要：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 discuss the feasibility of lipL32-PCR for detecting Leptospira interrogans. Methods The specificity and sensitivity of primers based on the lipL32 gene were compared with the recommended standard G1/G2 primers. Using these gene-based primers, a PCR assay was conducted for detection of L. interrogans in kidney samples from frogs and rats in Changshan county. PCR identification was performed on isolates taken from Zhejiang in 2008. Results The lipL32-PCR was specific and sensitive to L. interrogans. The PCR identification result for 2008 isolates was consistent with serological testing. A consistency of 95.0% was noted between lipL32-PCR and the assay based on the standard G1/G2 primer. The positive rate was 10.0% for lipL32-PCR and 5.0% for G1/G2-PCR. No significant difference between the two positive rates was shown by Fisher＇ s exact probability test （P：0.25）. Conclusion The lipL32-PCR was specific and sensitive in the detection of L. interrogans and can effectively reflect the carrier rate in animals, thus, providing evidence for the control of leptospirosis.

关 键 词：外膜脂蛋白基因 钩端螺旋体 聚合酶链反应 

分 类 号：R377.5[医药卫生—病原生物学]