一种新开发的用于检测马生殖道泰勒菌并与猪生殖道泰勒菌进行鉴别诊断的实时聚合酶链反应的评估。

Evaluation of a newly developed real-time PCR for the detection of Taylorella equigenitalis and discrimination from T. asinigenitalis.

作者信息

Premanandh J, George L V, Wernery U, Sasse J

机构信息

Central Veterinary Research Laboratory, P.O. Box 597, Dubai, United Arab Emirates.

出版信息

Vet Microbiol. 2003 Sep 24;95(4):229-37. doi: 10.1016/s0378-1135(03)00065-8.

DOI:10.1016/s0378-1135(03)00065-8

PMID:12935749

Abstract

A 'culture-LightCycler PCR' assay has been developed for the detection of Taylorella equigenitalis, the causative agent of contagious equine metritis (CEM) in horses. The primers and hybridisation probes were derived from the 16S rDNA sequence. Their specificity was determined in two closely related organisms and six commensal bacteria of the genital tract. The assay was specific for T. equigenitalis and discriminates T. asinigenitalis isolates. It also avoids misidentifications of morphologically and phenotypically similar organisms. The sensitivity was evaluated in comparison to a standard bacteriological culture method. It detected T. equigenitalis in 10 of 52 samples that had not been identified bacteriologically. The results indicated that the assay had a greater sensitivity. This is the first real-time PCR for the detection of T. equigenitalis and avoids PCR carry-over contamination. The 'culture-LightCycler PCR' assay is specific, sensitive and reproducible, and can be used effectively for the detection of T. equigenitalis infections.

摘要

已开发出一种“培养-罗氏LightCycler PCR”检测方法，用于检测马生殖道感染性子宫内膜炎（CEM）的病原体马泰勒氏菌。引物和杂交探针源自16S rDNA序列。在两种密切相关的生物体和六种生殖道共生细菌中确定了它们的特异性。该检测方法对马泰勒氏菌具有特异性，可区分亚洲泰勒氏菌分离株。它还避免了对形态和表型相似生物体的错误鉴定。与标准细菌培养方法相比，评估了该检测方法的灵敏度。在52份未通过细菌学鉴定的样本中，有10份检测到了马泰勒氏菌。结果表明该检测方法具有更高的灵敏度。这是首次用于检测马泰勒氏菌的实时PCR方法，可避免PCR产物污染。“培养-罗氏LightCycler PCR”检测方法具有特异性、灵敏性和可重复性，可有效用于检测马泰勒氏菌感染。