A novel polymerase chain reaction assay to detect Mycoplasma genitalium.

Molecular pathology : MP Pub Date : 2003-02-01 DOI:10.1136/mp.56.1.25

K Eastick, J P Leeming, E O Caul, P J Horner, M R Millar

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引用次数: 26

Abstract

Aims: To design and validate a polymerase chain reaction (PCR) assay targeting the 16S rRNA gene of Mycoplasma genitalium.

Methods: Primers were designed that were complementary to the 16S rRNA gene sequence of M genitalium. After optimisation of the reaction conditions, the PCR was tested against nine M genitalium strains, a dilution series of M genitalium DNA, and a panel of common microorganisms. The PCR was also challenged in parallel with a published assay against 54 urine specimens from men with urethritis.

Results: The expected 341 bp product was produced on amplification of material from all M genitalium strains and from none of the other microorganisms tested. The lower limit of detection was 50 genome copies. The new assay detected M genitalium DNA in nine of 54 men with urethritis, in comparison with eight positive specimens detected with the alternative PCR.

Conclusions: This novel PCR targeting the M genitalium 16S rRNA gene has been optimised and now provides a sensitive and specific alternative or addition to the available MgPa gene targeting assays.

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一种检测生殖支原体的聚合酶链反应新方法。

目的:设计并验证一种针对生殖器支原体16S rRNA基因的聚合酶链反应(PCR)检测方法。方法:设计与生殖支原体16S rRNA基因序列互补的引物。优化反应条件后，对9株生殖道分枝杆菌菌株、生殖道分枝杆菌DNA稀释系列和一组常见微生物进行PCR检测。聚合酶链反应也与一项已发表的针对54例尿道炎男性尿液标本的试验同时受到挑战。结果:所有生殖支原体菌株的扩增产物均为预期的341 bp，其他微生物均未扩增。检测下限为50个基因组拷贝。新的检测方法在54名尿道炎患者中检测到9名男性生殖器M DNA，而用另一种PCR检测到的阳性标本只有8名。结论:这种针对生殖器M - 16S rRNA基因的新型PCR已经过优化，现在为现有的MgPa基因靶向检测提供了一种敏感和特异性的替代或补充。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Molecular pathology : MP

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