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[结核分枝杆菌IS6110缺失株的聚合酶链反应检测]

[PCR detection of Mycobacterium tuberculosis lacking IS 6110].

作者信息

el Baghdadi J, Lazraq R, Benani A, Naciri M, Ibrahimy S, Benslimane A

机构信息

Unité des mycobactéries, Institut Pasteur du Maroc, Casablanca, Maroc.

出版信息

Bull Soc Pathol Exot. 1997;90(5):303-6.

PMID:9507757
Abstract

We have evaluated the frequency of M. tuberculosis strains which lack IS 6110 among 102 sputa isolated from Moroccan patients. A pair of primers was designed to amplify a 201bp DNA fragment of IS 6110. The amplified DNA was detected by ethidium bromide stained agarose gel electrophoresis and confirmed by southern blot hybridization with a 32P-labelled probe (PMTO2). To detect the presence of amplification inhibitors, an internal control DNA was added in each negative PCR result. Among 102 samples, 6 sputa were negative by PCR-IS 6110 but culture positive. The test of detection of M. tuberculosis for 2/6 sputa by PCR Amplicor amplifying 584 pb of rRNA 16s sequence was positive. RFLP analysis of these 2 strains revealed no bands hybridizing IS 6110 but PCR-Mt 308 was positive. These results confirmed that these M. tuberculosis strains are lacking IS 6110.

摘要

我们评估了从摩洛哥患者分离出的102份痰标本中缺乏IS 6110的结核分枝杆菌菌株的频率。设计了一对引物来扩增IS 6110的201bp DNA片段。扩增的DNA通过溴化乙锭染色的琼脂糖凝胶电泳进行检测,并用32P标记的探针(PMTO2)进行Southern印迹杂交确认。为了检测扩增抑制剂的存在,在每个阴性PCR结果中加入内部对照DNA。在102份样本中,6份痰标本经PCR-IS 6110检测为阴性,但培养结果为阳性。通过PCR Amplicor扩增16s rRNA的584 pb对2/6份痰标本进行结核分枝杆菌检测试验呈阳性。对这2株菌株的RFLP分析显示没有与IS 6110杂交的条带,但PCR-Mt 308呈阳性。这些结果证实这些结核分枝杆菌菌株缺乏IS 6110。

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