重复元件-聚合酶链反应用于临床和环境分离的军团菌属菌种的基因分型。

Repetitive element-polymerase chain reaction for genotyping of clinical and environmental isolates of Legionella spp.

机构信息

Department of Infectious, Respiratory, and Digestive Medicine, Control and Prevention of Infectious Diseases, Faculty of Medicine, University of the Ryukyus, Okinawa, Japan.

出版信息

Diagn Microbiol Infect Dis. 2010 Sep;68(1):7-12. doi: 10.1016/j.diagmicrobio.2010.04.001.

DOI:10.1016/j.diagmicrobio.2010.04.001

PMID:20727463

Abstract

Genotyping of Legionella strains is important for the epidemiologic survey of Legionnaires' disease infections. In this study, we investigated the potential of repetitive element-polymerase chain reaction (rep-PCR) for differentiating various isolates of Legionella spp. We used 38 Legionella pneumophila isolates (collected in clinics all over Japan between 1980 and 2007), 19 environmental Legionella anisa isolates (collected in Okinawa, Nara, Osaka, and Hyogo prefecture between 1987 and 2007), and 2 Legionella-type strains. We extracted bacterial genomic DNA and applied it to rep-PCR. PCR products were then converted into bands by agarose gel electrophoresis. The L. pneumophila serogroup (SG) 1 displayed very diverse patterns. Different bands were produced for each species of Legionella, and each species was clearly distinct. Phylogenetic analysis displayed 1 cluster of L. anisa isolates, while other Legionella spp. were present at discrete levels. Our findings show that rep-PCR is an effective, rapid, and simple technique for differentiation of L. pneumophila strains as well as Legionella spp.

摘要

对军团菌株进行基因分型对于军团病感染的流行病学调查非常重要。在这项研究中，我们研究了重复元件聚合酶链反应（rep-PCR）在区分各种军团菌属分离株方面的潜力。我们使用了 38 株嗜肺军团菌分离株（1980 年至 2007 年在日本各地诊所采集）、19 株环境嗜肺军团菌分离株（1987 年至 2007 年在冲绳、奈良、大阪和兵库县采集）和 2 株军团菌型菌株。我们提取了细菌基因组 DNA，并将其应用于 rep-PCR。然后通过琼脂糖凝胶电泳将 PCR 产物转化为带。嗜肺军团菌血清群（SG）1 显示出非常多样化的模式。每个军团菌属都产生了不同的条带，每个属都非常明显。系统发育分析显示出嗜肺军团菌分离株的 1 个聚类，而其他军团菌属则处于离散水平。我们的研究结果表明，rep-PCR 是一种有效、快速且简单的技术，可用于区分嗜肺军团菌菌株以及其他军团菌属。