Identification of macrolide-resistant Mycoplasma genitalium using real-time PCR
Identification of macrolide-resistant Mycoplasma genitalium using real-time PCR
复制标题
DOI：
10.1111/jdv.12963
复制
发表时间：
2015-08-01
影响因子：
9.2
通讯作者：
Reinton, N.
中科院分区：
文献类型：
--
作者：
Wold, C.;Sorthe, J.;Reinton, N.
生殖支原体是西欧非淋菌性尿道炎（NGU）的常见病因，但并非所有诊所都进行常规检测。据报道，大环内酯类耐药生殖支原体的患病率很高。一个易于使用的测试，可以预测可能的大环内酯类药物治疗失败是潜在的非常有价值的。我们报告的发展，一个快速，可靠的实时PCR检测生殖支原体23S rRNA基因的所有相关的耐药位点。MethodsMycoplasma genitalium阳性的临床样本收集在2012年12月和2013年5月之间，从样本常规发送到实验室进行生殖支原体诊断检测。实时PCR检测设计使用正向扩增引物互补的所有相关的共同确定的23 S rRNA基因突变，一个共同的反向扩增引物和一个共同的TaqMan Probe. ResulttWe报告Taqman检测常见的23 S rRNA基因型在位置2058和2059（大肠杆菌编号）与大环内酯类耐药，直接从临床样本。我们通过与DNA序列determination.ConclusionOur TaqMan检测方法检测与大环内酯类耐药生殖支原体，即A2058G，A2059G和A2058C相关的常见基因型。我们显示了耐药生殖支原体的存在与治疗失败之间的关联，从而证实了检测这些突变体的有效性，以防止抗菌素耐药性的进一步传播，并允许知情选择抗生素进行治疗。
ObjectivesMycoplasma genitalium is a common cause of non-gonococcal urethritis (NGU) in Western Europe, but is not routinely tested for in all clinics. A high prevalence of macrolide-resistant M.genitalium has been reported. An easy to use test that can predict likely macrolide treatment failure is potentially very valuable. We report the development of a rapid and reliable real-time PCR-assay which detects all relevant resistance loci in the M.genitalium 23S rRNA gene.MethodsMycoplasma genitalium-positive clinical samples were collected between December 2012 and May 2013, from samples sent routinely to the laboratory for diagnostic testing for M.genitalium. The real-time PCR assay was designed using forward amplification primers complementary to all relevant commonly identified 23s rRNA gene mutations, a common reverse amplification primer and a common TaqMan Probe.ResultWe report a Taqman assay for detection of common 23S rRNA genotypes at position 2058 and 2059 (Escherichia coli numbering) associated with macrolide resistance, directly from clinical samples. We validated the assay by comparison with DNA sequence determination.ConclusionOur TaqMan assay detects common genotypes associated with macrolide-resistant M.genitalium, namely, A2058G, A2059G and A2058C. We show association between the presence of resistant M.genitalium and treatment failure, thereby confirming the validity of testing for these mutants to prevent further spread of antimicrobial resistance and to allow informed choice of antibiotics for treatment.