基于聚合酶链反应（PCR）对胸膜肺炎放线杆菌生物变种I和II的2型分离株进行鉴定。

PCR-based identification of serotype 2 isolates of Actinobacillus pleuropneumoniae biovars I and II.

作者信息

Hüssy Daniela, Schlatter Yvonne, Miserez Raymond, Inzana Thomas, Frey Joachim

机构信息

Institute of Veterinary Bacteriology, University of Berne, Laenggass-Strasse 122, CH-3001 Bern, Switzerland.

出版信息

Vet Microbiol. 2004 Apr 19;99(3-4):307-10. doi: 10.1016/j.vetmic.2003.12.007.

DOI:10.1016/j.vetmic.2003.12.007

PMID:15066734

Abstract

A genetic typing method utilizing PCR for the identification of Actinobacillus pleuropneumoniae serotype 2 isolates has been developed based on the in vitro amplification of a 1.4 kb DNA segment of the serotype 2 capsular polysaccharide genes cps2AB. The assay was tested with all serotype reference strains and a collection of 92 different A. pleuropneumoniae strains of all 15 serotypes of both biovars I and II, originating from 18 different countries worldwide. The cps2 based PCR identified the serotype 2 reference strain and all 12 serotype 2 collection strains contained in this set. DNA was not amplified from the remaining A. pleuropneumoniae reference and collection strains, indicating the PCR assay was highly specific. Furthermore, the PCR method detected all 31 A. pleuropneumoniae serotype 2 field isolates from diseased pigs that were identified in parallel as serotype 2 by agar gel diffusion. The serotype 2 PCR assay proved to be highly specific and reliable for the identification of serotype 2 isolates of A. pleuropneumoniae.

摘要

一种利用聚合酶链反应（PCR）鉴定胸膜肺炎放线杆菌2型分离株的基因分型方法已经建立，该方法基于对2型荚膜多糖基因cps2AB的1.4 kb DNA片段进行体外扩增。该检测方法用所有血清型参考菌株以及来自全球18个不同国家的92株不同的胸膜肺炎放线杆菌菌株进行了测试，这些菌株涵盖了生物变种I和II的所有15种血清型。基于cps2的PCR鉴定出了该组中的2型参考菌株以及所有12株2型收集菌株。未从其余的胸膜肺炎放线杆菌参考菌株和收集菌株中扩增出DNA，这表明该PCR检测具有高度特异性。此外，该PCR方法检测出了来自患病猪的所有31株胸膜肺炎放线杆菌2型田间分离株，这些分离株通过琼脂凝胶扩散法同时被鉴定为2型。事实证明，2型PCR检测方法对于鉴定胸膜肺炎放线杆菌2型分离株具有高度特异性和可靠性。