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目的建立沙眼衣原体(Ct)主要外膜蛋白基因(omp1)上游引物5′末端地高辛标记裂解酶片段长度多态性(CFLP)分型方法,并对Ct临床分离株进行分型。方法取临产孕妇宫颈刮片及其新生儿鼻咽拭子组成母婴配对标本300对605份,用套式聚合酶链反应扩增Ctomp1第四可变区;用建立的CFLP方法对其进行基因分型。结果孕妇宫颈Ct检出率11%(33/300),母婴传播率24.2%(8/33);8对Ct阳性母婴配对标本CFLP图谱呈四类,经测序证实分别为E、F、H、D型(各3、2、2、1对),各占37.5%、25%、25%和12.5%,且每对母子CFLP图谱完全一致;同一标本不同批次和相同基因型不同标本的CFLP图谱一致。结论上游引物5′末端地高辛标记Ctomp1CFLP分型方法灵敏度高、重复性好、操作简便、快速、费用相对低廉,无放射污染,是一种极具潜力的Ct基因分型方法。
OBJECTIVE: To establish a method for genotyping Ct clinical isolates by digoxigenin-labeled fragment length polymorphism (PCR-RFLP) at the 5 ’end of the upstream primer of the major outer membrane protein gene (omp1) of Chlamydia trachomatis (Ct). Methods Three hundred and fifty-five pairs of maternal and neonatal paired specimens were obtained from cervical smears of pregnant women and nasopharyngeal swabs of neonates. The fourth variable region of Ctomp1 was amplified by nested polymerase chain reaction (PCR). The CFLP gene Type. Results The detection rate of cervical Ct in pregnant women was 11% (33/300) and the rate of mother to child transmission was 24.2% (8/33). There were four types of CFLP maps in 8 paired samples of Ct positive maternal and infant, which were confirmed as E, F, H, D type (each 3,2,2,1 pairs), each accounting for 37.5%, 25%, 25% and 12.5%, and each mother and child exactly the same CFLP pattern; the same specimen different batches and the same genotype different specimens The CFLP pattern is consistent. Conclusion The method of Ctomp1CFLP on the 5’-end of the upstream primer is highly sensitive and reproducible. It is simple, rapid, inexpensive, and free of radioactive contamination. It is a potential Ct genotyping method.