应用PCR技术检测致病性钩端螺旋体  

Detection of all serogroups of pathogenic leptospires epidemic by PCR

作　　者：唐雨德[1] 周东明[1] 陈永红[1] 李越希[1] 陶开华[1] 

机构地区：[1]南京军区军事医学研究所军队卫生学研究室,南京210002

出　　处：《中国公共卫生》2004年第9期1042-1044,共3页Chinese Journal of Public Health

基　　金：全军"十五"医学计划资助项目 (0 1L0 0 6)

摘　　要：目的　建立一种能特异检测我国致病性钩端螺旋体所有血清型的PCR方法。方法　从Genbank中选取钩体 2 3SrDNA序列 ,设计一对种特异性引物。通过Blast验证引物的特异性和广谱性后 ,用PCR检测我国流行的所有 18个血清群钩体代表株 ,观察其敏感性和特异性。探讨简便的样品处理方法 ,并对模拟标本进行检测。结果　18个血清群的 2 5株钩体均出现单一 4 82bp的特异性扩增产物 ,而双曲钩体及其他螺旋体、微生物、空白对照均无任何DNA扩增条带。PCR单一反应最小检出钩体数为 8条。用煮沸法、试剂盒、SiO高盐吸附法及氯仿苯酚混合法处理样品对PCR结果无明显影响。对模拟标本的检测符合临床实际要求。结论　PCR方法是一种灵敏度高、特异性强、检测谱广的快速检测钩体的有效方法。Objective To develop a polymerase chain reaction (PCR) for specifically detecting all serogroups of leptospira interrogans epidemic in China.Methods A pair of species-specific primers were designed from an 23s rDNA sequence of leptospira interrogans from Genbank.After the primers were identified they were specific and universal by Blast,18 serogroups of Leptospira interrogans and other pathohens or microbes were detected by PCR.Its sensitivity and specifity were tested.The different sample preparating method was developed,and sera and water samples were tested.Results A specific amplification product with 482?bp length which could be found from eighteen serogroups of leptospira interrogans by PCR,and agarose gel electrophoresis was demonstrated to have a sensitivity of detecting DNAs of eight L.Interrogans.None of DNA amplification products was found from other microbes.Conclusion PCR was a rapid,specific and sensitive and universal method for detecting leptospira in the epidemic regions.

关 键 词：致病性钩端螺旋体 PCR 血清群 

分 类 号：R514[医药卫生—内科学]