一种用于检测斑点热群和斑疹伤寒群立克次体的高灵敏度和特异性实时PCR检测方法。

A highly sensitive and specific real-time PCR assay for the detection of spotted fever and typhus group Rickettsiae.

作者信息

Stenos John, Graves Stephen R, Unsworth Nathan B

机构信息

The Australian Rickettsial Reference Laboratory, Department of Clinical and Biomedical Sciences, The University of Melbourne, The Geelong Hospital, Victoria, Australia.

出版信息

Am J Trop Med Hyg. 2005 Dec;73(6):1083-5.

PMID:16354816

Abstract

A highly specific real-time polymerase chain reaction (PCR) assay was developed to detect spotted fever and typhus group rickettsiae using the citrate synthase gene as the target. The assay amplified rickettsial members of the spotted fever and typhus group including Rickettsia akari, R. australis, R. conorii, R. honei, "R. marmionii," R. sibirica, R. rickettsii, R. typhi, and R. prowazekii. The ancestral group rickettsia, R. bellii, did not produce a positive reaction, nor did other members of the order Rickettsiales or any non-rickettsial bacteria. The assay had a sensitivity of one target copy number per reaction as determined by serial dilutions of a plasmid containing a spotted fever group target sequence. This quantitative assay is useful for the enumeration of rickettsiae in clinical specimens and the diagnosis of rickettsial illnesses, when rickettsial numbers are very low.

摘要

开发了一种高度特异性的实时聚合酶链反应（PCR）检测方法，以柠檬酸合酶基因作为靶标来检测斑点热群和斑疹伤寒群立克次体。该检测方法可扩增斑点热群和斑疹伤寒群的立克次体成员，包括小蛛立克次体、澳大利亚立克次体、康氏立克次体、霍氏立克次体、“马氏立克次体”、西伯利亚立克次体、立氏立克次体、伤寒立克次体和普氏立克次体。原始群立克次体——贝利立克次体未产生阳性反应，立克次体目其他成员或任何非立克次体细菌也未产生阳性反应。通过对含有斑点热群靶序列的质粒进行系列稀释测定，该检测方法的灵敏度为每个反应一个靶标拷贝数。当立克次体数量非常低时，这种定量检测方法对于临床标本中立克次体的计数和立克次体病的诊断很有用。