High-throughput multistrain polymerase chain reaction quantification of Chlamydia trachomatis from clinical and preclinical urogenital specimens
High-throughput multistrain polymerase chain reaction quantification of Chlamydia trachomatis from clinical and preclinical urogenital specimens
复制标题
DOI：
10.1016/j.diagmicrobio.2009.01.032
复制
发表时间：
2009-06-01
影响因子：
2.9
通讯作者：
Pyles, Richard B.
中科院分区：
文献类型：
--
作者：
McGowin, Chris L.;Whitlock, Gregory C.;Pyles, Richard B.
沙眼衣原体（CT）是世界上最普遍的性传播细菌病原体，可引起严重的生殖道感染。目前，核酸扩增试验（NAATs）是临床诊断的金标准，但大多数NAATs是劳动密集型的，仅限于特定的CT血清型。我们开发并验证了一种定量聚合酶链反应（qPCR）检测方法，该方法可在2 log（10）到10 log（10）个基因组的线性范围内重复性地检测CT血清型D、E、F、la和muridarum衣原体，并且从实验和人类尿液样本中获得低变异系数。人类阴道、宫颈内和男性尿道拭子的CT DNA负荷与BD ProbeTec ET测定（Becton Dickinson诊断系统，Franklin Lakes， NJ）并行运行具有良好的相关性。在一项临床前杀微生物剂评估中，小鼠拭子和组织中的muridarum DNA负荷与免疫荧光测定具有良好的相关性。优化后的qPCR系统提高了灵敏度，便于临床和实验临床前样品的定量评价，用于抗ct治疗和杀微生物剂评价。(C) 2009爱思唯尔公司版权所有。
Chlamydia trachomatis (CT) is the most prevalent sexually transmitted bacterial pathogen worldwide and causes severe reproductive tract infections. Currently, nucleic acid amplification tests (NAATs) are the gold standard for clinical diagnosis, but most NAATs are labor intensive and limited to specific CT serovars. We developed and validated a quantitative polymerase chain reaction (qPCR) assay that reproducibly detected CT serovars D, E, F, la, and Chlamydia muridarum over a linear range of 2 log(10) to 10 log(10) genomes with low coefficients of variation from both experimental and human urine samples. CT DNA loads from human vaginal, endocervical, and male urethral swabs correlated well with the BD ProbeTec ET assay (Becton Dickinson Diagnostic Systems, Franklin Lakes, NJ) run in parallel. In a preclinical microbicide evaluation, C. muridarum DNA loads in mouse swabs and tissues correlated well with an immunofluorescence assay. The optimized qPCR system provided enhanced sensitivity and facilitated the quantitative evaluation of clinical and experimental preclinical samples for anti-CT therapeutic and microbicide evaluation. (C) 2009 Elsevier Inc. All rights reserved.