聚合酶链反应与限制性内切酶分析联合应用鉴别衣原体种类

Differentiation of Chlamydia species by combined use of polymerase chain reaction and restriction endonuclease analysis.

作者信息

Yoshida H, Kishi Y, Shiga S, Hagiwara T

机构信息

Department of Virology I, National Institute of Infectious Diseases, Tokyo, Japan.

出版信息

Microbiol Immunol. 1998;42(5):411-4. doi: 10.1111/j.1348-0421.1998.tb02303.x.

DOI:10.1111/j.1348-0421.1998.tb02303.x

PMID:9654375

Abstract

To differentiate Chlamydia spp., a primer pair designed to generate a genus-specific region of the major outer membrane protein (MOMP) gene was used in a PCR to amplify a single DNA fragment of 245-259 bp. In the PCR, the expected single DNA fragment was amplified from strains of Chlamydia trachomatis, C. psittaci, C. pneumoniae and C. pecorum, respectively. By restriction endonuclease analysis with AluI and PvuII, the amplified products exhibited four distinct patterns, corresponding to the four species. It is, therefore, concluded that one-step PCR followed by restriction endonuclease analysis as described in this study could be a valuable method for the detection and differentiation of Chlamydia species.

摘要

为了区分衣原体属物种，设计了一对引物用于扩增主要外膜蛋白（MOMP）基因的属特异性区域，该引物对用于聚合酶链反应（PCR）以扩增出一条245 - 259 bp的单一DNA片段。在PCR反应中，分别从沙眼衣原体、鹦鹉热衣原体、肺炎衣原体和猪衣原体的菌株中扩增出了预期的单一DNA片段。通过用AluI和PvuII进行限制性内切酶分析，扩增产物呈现出四种不同的模式，分别对应这四个物种。因此，可以得出结论，如本研究中所述的一步PCR随后进行限制性内切酶分析可能是一种用于检测和区分衣原体物种的有价值的方法。