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应用聚合酶链反应检测结核分枝杆菌复合群DNA以快速诊断皮肤结核。

Detection of Mycobacterium tuberculosis complex DNA by the polymerase chain reaction for rapid diagnosis of cutaneous tuberculosis.

作者信息

Margall N, Baselga E, Coll P, Barnadas M A, de Moragas J M, Prats G

机构信息

Department of Microbiology, Universitat Autònoma de Barcelona, Spain.

出版信息

Br J Dermatol. 1996 Aug;135(2):231-6.

PMID:8881665
Abstract

We assessed the polymerase chain reaction (PCR) technique to detect Mycobacterium tuberculosis complex DNA in 48 paraffin-embedded specimens from 32 patients with different variants of cutaneous tuberculosis, and compared the results with those of culture. A 123 bp product of the IS6110 insertion sequence specific of M. tuberculosis complex was amplified and confirmed by digestion with SalI restriction endonuclease. The time required for the procedure was 3 days. Thirty-seven samples (77.1%) were positive for M. tuberculosis complex DNA. No false positive results were obtained in nine negative controls. Of the 20 specimens tested by PCR and culture, the frequency of positivity was 90% for DNA amplification and 65% for culture. In seven cases of lupus vulgaris, the figures were 100% and 57%, respectively. In the 11 specimens culture negative or not microbiologically tested and PCR negative, evidence for tuberculous infection was provided by the correlation of various relative and absolute criteria. These results show that PCR amplification of the IS6110 insertion fragment is a rapid and accurate means for the detection of M. tuberculosis complex DNA in paraffin-embedded skin biopsies from patients with cutaneous tuberculosis, especially in paucibacillary lesions.

摘要

我们评估了聚合酶链反应(PCR)技术,以检测32例患有不同类型皮肤结核患者的48份石蜡包埋标本中的结核分枝杆菌复合群DNA,并将结果与培养结果进行比较。扩增出结核分枝杆菌复合群特异性的IS6110插入序列的123 bp产物,并用SalI限制性内切酶消化进行确认。该过程所需时间为3天。37个样本(77.1%)的结核分枝杆菌复合群DNA呈阳性。9个阴性对照未获得假阳性结果。在20份同时进行PCR和培养检测的标本中,DNA扩增的阳性率为90%,培养的阳性率为65%。在7例寻常狼疮病例中,这两个数字分别为100%和57%。在11份培养阴性或未进行微生物检测且PCR阴性的标本中,通过各种相对和绝对标准的相关性提供了结核感染的证据。这些结果表明,IS6110插入片段的PCR扩增是检测皮肤结核患者石蜡包埋皮肤活检中结核分枝杆菌复合群DNA的快速、准确方法,尤其是在少菌性病变中。

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