摘要
对检测麻风菌用的几种PCR方法进行了比较研究。这些PCR以编码麻风菌65KD、36KD、18KD、groEL和16S rRNA的基因为基础。比较用的指标是敏感性、特异性、简易快速和节省试剂,结果显示以编码麻风菌65KD抗原的基因为基础的PCR(此法原为Woods报告,后经我们优化改良)为最实用,其检测下限为10～100条麻风菌,与其它已知的分枝菌无交叉反应,用单对引物,仅需30个循环,以2.5～3个小时就能完成实验,且用料仅为其它常用反应体系的1/2,敏感性高、特异性强,简单、快速、价廉,故适于推广应用。
A study on comparison between several PCRs for detection of M. leprae was done. The PCRs were established on the basis of M. leprae genes coding 65KD, 36KD, 18KD, groEL antigens and 16s rRNA.. The indicators for the comparison are sensitivity, specificity,simplicity, spee ,economic cost and so on. The results indicated that the PCR with M. leprae gene coding 65KD antigen, which was developed by Woods et al and then modified by Wu et al ,was of choice. The lowest limit of its detection is 10 to 100 AFBs, there was no crossreaction with other mycobacteria used in the test, and it only needs one pair of primers. The optimum number of cycles are 30 which consume 2. 5 to 3 hrs, and the volume of reaction system is 50 μl which save 50% of the materials in comparison with other PCRs. The authors consider that this PCR is the most valuable to be popularized for detection of M. leprea.
基金
国家自然科学基金
