Femeris Women's Health Research Center, Medical Diagnostic Laboratories, L.L.C., A Member of Genesis Biotechnology Group, Hamilton, NJ.
Femeris Women's Health Research Center, Medical Diagnostic Laboratories, L.L.C., A Member of Genesis Biotechnology Group, Hamilton, NJ.
Diagn Microbiol Infect Dis. 2014 Apr;78(4):321-7. doi: 10.1016/j.diagmicrobio.2013.08.004. Epub 2014 Jan 18.
Lactobacilli play a key role in promoting vaginal health. Depletion of these bacteria is associated with bacterial vaginosis (BV), the most common vaginal disorder. Here we describe the development and laboratory validation of a novel single-tube multiplex TaqMan quantitative polymerase chain reaction (qPCR) assay for the identification and quantitative assessment of the four major vaginal Lactobacillus species: L. crispatus, L. jensenii, L. gasseri, and L. iners. The assay utility was evaluated by the analysis of lactobacilli in non-cultured clinical vaginal swab specimens collected from BV patients and healthy individuals. As confirmed by the assay, L. crispatus, L. jensenii, and to a lesser extent L. gasseri, are common in the vagina of healthy women, whereas L. iners dominance is associated with BV. The major assay limitation was preferential detection of dominant Lactobacillus species in samples with mixed lactobacilli resulting in lower sensitivity for minor species. The multiplex qPCR assay described here is an advance in the detection and quantitation of the major vaginal lactobacilli, potentially facilitating the molecular diagnosis of BV and post-therapy restoration of the vaginal microflora.
乳杆菌在促进阴道健康方面发挥着关键作用。这些细菌的消耗与细菌性阴道病（BV）有关，BV 是最常见的阴道疾病。在这里，我们描述了一种新型的单管多重 TaqMan 实时聚合酶链反应（qPCR）检测方法的开发和实验室验证，用于鉴定和定量评估四种主要阴道乳杆菌：卷曲乳杆菌、詹氏乳杆菌、加氏乳杆菌和惰性乳杆菌。通过分析来自 BV 患者和健康个体的非培养临床阴道拭子标本，评估了该检测方法的实用性。该检测方法证实，卷曲乳杆菌、詹氏乳杆菌和在较小程度上的加氏乳杆菌在健康女性的阴道中很常见，而惰性乳杆菌的优势与 BV 有关。该检测方法的主要局限性是在混合乳杆菌的样本中优先检测优势乳杆菌物种，从而降低了对次要物种的检测灵敏度。这里描述的多重 qPCR 检测方法是检测和定量主要阴道乳杆菌的一项进展，可能有助于 BV 的分子诊断和治疗后阴道微生物群的恢复。
