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使用聚合酶链反应(PCR)检测牛奶中的伯氏考克斯氏体。

Detection of Coxiella burnetii in cow's milk using the polymerase chain reaction (PCR).

作者信息

Willems H, Thiele D, Frölich-Ritter R, Krauss H

机构信息

Institut für Hygiene und Infektionskrankheiten der Tiere, Justus-Liebig-Universität, Giessen, Germany.

出版信息

Zentralbl Veterinarmed B. 1994 Nov;41(9):580-7. doi: 10.1111/j.1439-0450.1994.tb00267.x.

Abstract

A PCR approach (transposon PCR) with primers based on repetitive transposon-like sequences, which--depending on the isolate--were found at a minimum frequency of 19 on the C. burnetii genome, was established for the highly sensitive and specific detection of C. burnetii. This study describes the analytical detection of C. burnetii in milk, which requires a special preparation method prior to PCR. Because of the low level of C. burnetii particles in milk samples, template DNA was concentrated by a factor of 200, using cetyltrimethylammonium bromide as the precipitation reagent. Using this particular preparation method, even a single C. burnetii particle could be detected in 1 ml milk.

摘要

建立了一种基于重复转座子样序列引物的PCR方法(转座子PCR),根据分离株的不同,这些序列在伯氏考克斯氏体基因组上的出现频率至少为19次,用于伯氏考克斯氏体的高灵敏度和特异性检测。本研究描述了牛奶中伯氏考克斯氏体的分析检测,在进行PCR之前需要一种特殊的制备方法。由于牛奶样品中伯氏考克斯氏体颗粒水平较低,使用十六烷基三甲基溴化铵作为沉淀剂,将模板DNA浓缩了200倍。使用这种特殊的制备方法,在1毫升牛奶中甚至可以检测到单个伯氏考克斯氏体颗粒。

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