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利用聚合酶链反应检测和区分伯氏考克斯氏体菌株的策略。

Strategy for detection and differentiation of Coxiella burnetii strains using the polymerase chain reaction.

作者信息

Mallavia L P, Whiting L L, Minnick M F, Heinzen R, Reschke D, Foreman M, Baca O G, Frazier M E

机构信息

Washington State University, Pullman 99164.

出版信息

Ann N Y Acad Sci. 1990;590:572-81. doi: 10.1111/j.1749-6632.1990.tb42268.x.

Abstract

A method for the rapid detection of Coxiella burnetii and differentiation between strains that cause endocarditis and those that cause acute Q fever is based on the observation that the different strains contain unique plasmid sequences. This method employs the polymerase chain reaction (PCR) and requires knowledge of specific DNA sequences in the region (target) of DNA to be amplified. To detect and differentiate between C. burnetii isolates, two sets of primers are required. The first set was derived from a fragment of plasmid QpH1 which has been detected in all C. burnetii isolates. A second PCR reaction was conducted using primers specific for DNA sequences that are shared only by QpRS plasmid-containing strains of C. burnetii. The first reaction detects the presence of C. burnetii. The second PCR is necessary to determine whether the isolate contains DNA sequences associated with strains causing chronic disease. These procedures detect as few as one to ten organisms.

摘要

一种快速检测伯氏考克斯体以及区分引起心内膜炎的菌株和引起急性Q热的菌株的方法,是基于不同菌株含有独特质粒序列这一观察结果。该方法采用聚合酶链反应(PCR),并且需要了解待扩增DNA区域(靶标)中的特定DNA序列。为了检测和区分伯氏考克斯体分离株,需要两组引物。第一组引物源自质粒QpH1的一个片段,该片段在所有伯氏考克斯体分离株中均被检测到。使用仅对含有QpRS质粒的伯氏考克斯体菌株共有的DNA序列特异的引物进行第二次PCR反应。第一次反应检测伯氏考克斯体的存在。第二次PCR对于确定分离株是否含有与引起慢性病的菌株相关的DNA序列是必要的。这些程序能够检测低至一到十个生物体。

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