聚合酶链反应(PCR)检测水中钩体DNA方法的建立和初步应用

ESTABLISHMENT AND PRELIMINARY APPLICATION OF DETECTON OF LEP-TOSPIRAL DNA BY POLYMERASE CHAIN REACTION

目的:为了建立一个检测水中钩体DNA的技术方法。方法:本研究采用了根据1993年C.Gravekamp设计的引物G_1G_2序列自行合成的引物G_1G_2建立了检测致病钩体DNA的PCR方法。运用此方法并结合“优筛法”对采自云南河口、蒙自等地区108份水样进行检测,并与分离培养后经中国药品生物制品检定所鉴定结果进行了比较。结果:两组方法检测所有水样呈阴性,其精确度和一致性达100％,但分离培养方法漏检率为8.3％(9/108)。结论:建立了PCR检测水中钩体DNA方法。此方法简单、快速、准确,适用于一般实验室及大规模水样钩体DNA的检测,值得推广应用。

Objective: In order to establish a sensitive method to detect leptospiral DNA in water. Method: With primer G, and G2on the basis of nucleotide sequence developed by Gravekamp in 1993, it was developed a polymerase chain reaction (PCR) technique detecting leptospiral DNA in water. 108 water samples from Menzhi and Hekou of Yunnan province were detected, and then the results were compared with those identified by National Institute for Control of Phamaceutical and Biological Products after cultivation. Results: All detected water samples were negative. Both accuracy and same rate are 100%, but the missed rate of cultiviation examination was 8. 3%. Conclusion:This method is simple, rapid and accurate. It is sui table for the routine detection of leptospiral DNA in laboratories and large -scale water investigation.