Identification and Quantification of Waddlia Chondrophila in Women with Recurrent Pregnancy Loss

Saeed Harirzadeh, F. Kafilzadeh, H. Zandi, M. Kargar
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Abstract

Background and Aims: Recurrent pregnancy loss (RPL) is one of the important complications of pregnancy. Only 50% of cases have a known cause of RPL. One of the causes of RPL is Waddlia Chondrophila (W. Chondrophila), but no information is available regarding its in Iran. This study aimed to develop a Taqman Real-Time polymerase chain reaction assay to detect recA Gene of W. Chondrophilain biological samples from women with RPL in Yazd city. Materials and Methods: Clinical samples of women with a history of RPL, normal childbirth, and standard strain WSU 86–1044 were provided. Primers and probes of W. Chondrophila were designed. Positive control was provided based on the PUC57 vector. Technical performance was examined using a control plasmid. Results: Analytical sensitivity was 5 ng/µl of control plasmid DNA. No cross-amplification was observed when testing other pathogens that can detect in human vaginitis. The Taqman Real-Time PCR assay exhibited good reproducibility. This Real-Time PCR was then applied to 100 vaginal swabs. No samples were revealed to be Waddlia positive. Conclusions: This new TaqMan Real-Time PCR assay represents a diagnostic tool that may be used to further study the prevalence of W. Chondrophila infection in clinical specimens.
复发性流产妇女Waddlia软骨病的鉴定和定量
背景与目的:复发性妊娠丢失(RPL)是妊娠的重要并发症之一。只有50%的病例有已知的RPL病因。RPL的原因之一是Waddlia软骨病(W. Chondrophila),但在伊朗没有关于其的信息。本研究旨在建立Taqman实时聚合酶链反应法检测亚兹德市RPL妇女嗜软骨杆菌生物样品的recA基因。材料和方法:提供有RPL病史,正常分娩的妇女临床样本,标准菌株WSU 86-1044。设计了软骨菌的引物和探针。以PUC57载体为阳性对照。用对照质粒检测技术性能。结果:对照质粒DNA的分析灵敏度为5 ng/µl。在检测人类阴道炎中可检测到的其他病原体时,未观察到交叉扩增。Taqman Real-Time PCR重现性好。然后将Real-Time PCR应用于100个阴道拭子。没有样本显示Waddlia呈阳性。结论:这种新的TaqMan Real-Time PCR检测是一种诊断工具,可用于进一步研究临床标本中嗜软骨芽胞杆菌感染的患病率。
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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