中华皮肤科杂志 ›› 2000, Vol. 33 ›› Issue (6): 401-403.

• 论著 • 上一篇    下一篇

多重聚合酶链反应用于D-K型沙眼衣原体基因分型的研究

刘全中, 陈锦英, 缴稳苓, 田静群, 傅志宜   

  1. 天津性传播疾病研究所 300052
  • 收稿日期:1999-12-09 出版日期:2000-12-15 发布日期:2000-12-15

Genotyping of Chlamydia trachomatis Isolates by a Multiplex PCR

LIU Quanzhong, CHEN Jinying, JIAO Wenling   

  1. Tianjin Institute of Sexually Transmitted Diseases. Tianjin 300052
  • Received:1999-12-09 Online:2000-12-15 Published:2000-12-15

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摘要： 目的 摸索一套泌尿生殖道沙眼衣原体基因分型的多重聚合酶链反应法。方法 选择某一型与其它各型均不同的可变区部位作为下游引物,以恒定区1区的高度保守区作为各型的公用上游引物,优化反应条件,进行标准株和临床野生株的多重聚合酶链反应扩增,并与聚合酶链反应-限制性内切酶片段多态性方法比较。结果 用摸索出的反应系统和条件,对8个标准株进行扩增,扩增出与理论值一致的片段,并可直接在普通水平琼脂糖凝胶进行电泳。特异性检测证明:①衣原体各型之间没有非特异性交叉扩增,②对泌尿生殖道病原和寄生微生物进行扩增,未扩出任何DNA带。对临床野生株分型结果证明:复合聚合酶链反应阳性率为100%,而聚合酶链反应-限制性内切酶片段多态性为94.44%。结论 本套衣原体基因分型的多重聚合酶链反应法特异、敏感、快速、实用,最终能用于临床分型。

关键词: 衣原体, 沙眼, 聚合酶链反应, 基因型

Abstract: Objective To establish a multiplex PCR genotyping method for Chlamydia trachomatis isolates. Methods With the aid of the special gene software, the homogenicity of C.trachomatis D-K omp1 gene was analysed, and the primers for genotyping were designed accordingly. The differences of amplified products of each genotype were over 80 bp so that they could be distinguished on agarose gel electrophoresis. The multiplex PCR was applied to test the reference and wild strains and was compared with PCR-RFLP method. Results After repeated experiments, each reference strain was amplified to reveal a clear band by the multiplex PCR which could be differentiated with each other on agarose gel electrophoresis. All wild strains could be genotyped by the multiplex PCR while 94.44% of the strains could be genotyped by PCR-RFLP. Conclusion The multiplex PCR genotyping method for C.trachomatis is a sensitive, specific, convenient and practical tool for clinical application.

Key words: Polymerase chain reaction, Genotype, Chlamydia trachomatis