PCR/RFLP对临床标本沙眼衣原体直接基因分型

建立泌尿生殖系统沙眼衣原体感染阳性标本直接基因分型方法。先用敏感的扩增沙眼衣原体特异质粒的引物扩增５００份临床标本，初筛选出１００份阳性标本再用扩增沙眼衣原体主要外膜蛋白（ＭＯＭＰ）基因的引物扩增，并用嵌套式ＰＣＲ使阳性标本能扩增出ＭＯＭＰ基因，对获得的ＭＯＭＰ基因用限制性酶切片段长度多态性（ＲＦＬＰ）方法进行基因分型，并用银染观察结果。其中Ｅ型比例最高，占４７％；Ｆ型为１６％；Ｄａ、Ｇ型均为６％；Ｂａ、Ｄ型各占５％；Ｈ型及Ｋ型均为１％，混合型Ｋ／Ｆ型２％；混合型Ｄ／Ｊ、Ｅ／Ｇ均为１％；未能分型有９％。

DIRECT GENOTYPING OF CHLAMYDIA TRACHOMATIS IN CLINICAL SAMPLES BY PCR RFLP METHODS

Five hundred clinical samples from urogenital infection patients were simply pretreated by boiling for 10 min and screened for chlamydia trachomatis plasmid PCR.One hundred positive samples tested by plasmid PCR were subjected to PCR for Omp 1 gene amplification,seventy eight samples were found positive by direct Omp 1 PCR while 22 direct Omp 1 negative samples became positive by the nested Omp 1 PCR.The amplified Omp 1 genes were treated with restriction enzymes Alu I, Hinf I, Dde I and Cfo I and the digested fragments were run on polyacrylamide gel with silver staining.Genotypes of Chlamydia trachomatis in the 100 clinical samples were identified by analysis of their restriction fragment length polymorphisms.The results showed that the genotype E was the most prevalent types found in the urogenital infection patients in our study,accounting for 47%(47/100)and the next prevalence was the genotype F 16%(16/100).Both genotypes Da and G made up 6%(6/100),both genotype Ba and D were 5%(5/100),and genotype H and K each was 1%(1/100).Mixed genotypes K/F were found in 2% and mixed genotypes D/J and E/G occurred in 1%.Nine samples were not identifiable because their RFLP patterns did not correspond to that of any of the 18 C.Trachomatis serovars tested in this study.