Targeted PCR for detection of vaginal bacteria associated with bacterial vaginosis
Targeted PCR for detection of vaginal bacteria associated with bacterial vaginosis
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DOI：
10.1128/jcm.01272-07
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发表时间：
2007-10-01
影响因子：
9.4
通讯作者：
Marrazzo, Jeanne M.
中科院分区：
文献类型：
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作者：
Fredricks, David N.;Fiedler, Tina L.;Marrazzo, Jeanne M.
在细菌性阴道病（BV）患者中检测到几种新型细菌，但这种方法对检测少数细菌不敏感。我们开发了一系列针对分类群的16S rRNA基因PCR检测方法，以更灵敏地检测阴道关键细菌。我们试图确定每种细菌在阴道中的流行程度，它们与细菌性阴道炎的关系，以及PCR在细菌性阴道炎微生物学诊断中的应用。建立了17种阴道细菌的靶向PCR检测方法，并应用于81名BV患者和183名非BV患者的264份阴道液样本。将结果与两种被广泛接受的诊断细菌性阴道炎的方法进行比较，即使用临床表现（Amsel标准）和解释阴道液革兰氏染色（Nugent标准）。细毛菌/Sneathia、阴道托波菌、一种蛋菌样细菌、Megasphaera种和三种梭菌目新细菌是与BV显著相关的细菌种类。与Amsel临床标准相比，PCR检测Megasphaera种或Clostridiales中的一种诊断BV的灵敏度为99%，特异性为89%；与Nugent标准（革兰氏染色）相比，灵敏度为95.9%，特异性为93.7%。PCR检测一种或多种挑剔的细菌种类是比检测细菌（如阴道加德纳菌）更可靠的BV指标，这突出了PCR诊断BV的潜力。
Several novel bacterial species have been detected in subjects with bacterial vaginosis (BV) by using broad-range PCR assays, but this approach is insensitive for detecting minority species. We developed a series of taxon-directed 16S rRNA gene PCR assays for more sensitive detection of key vaginal bacteria. We sought to determine the prevalence of each species in the vagina, its association with BV, and the utility of PCR for the microbiological diagnosis of BV. Targeted PCR assays were developed for 17 vaginal bacterial species and applied to 264 vaginal-fluid samples from 81 subjects with and 183 subjects without BV. The results were compared to those of two widely accepted methods for diagnosing BV, the use of clinical findings (Amsel criteria) and the interpretation of vaginal-fluid Gram stains (Nugent criteria). Leptotrichia/Sneathia, Atopobium vaginae, an Eggerthella-like bacterium, Megasphaera species, and three novel bacteria in the order Clostridiales are among the bacterial species significantly associated with BV. PCR detection of either a Megasphaera species or one of the Clostridiales bacteria yielded a sensitivity of 99% and a specificity of 89% for diagnosis of BV compared to the Amsel clinical criteria and a sensitivity of 95.9% and a specificity of 93.7% compared to the Nugent criteria (Gram stain). PCR detection of one or more fastidious bacterial species is a more reliable indicator of BV than detection of bacteria, such as Gardnerella vaginalis, previously linked to BV, highlighting the potential of PCR for the diagnosis of BV.