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

Saeed  Harirzadeh; Farshid  Kafilzadeh; Hengameh  Zandi; Mohammad Kargar

doi:10.18502/ijml.v9i4.11622

Saeed Harirzadeh Department of Biology, Jahrom Branch, Islamic Azad University, Jahrom, Iran
Farshid Kafilzadeh Department of Biology, Jahrom Branch, Islamic Azad University, Jahrom, Iran
Hengameh Zandi Department of Microbiology, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran
Mohammad Kargar Department of Biology, Jahrom Branch, Islamic Azad University, Jahrom, Iran

DOI: https://doi.org/10.18502/ijml.v9i4.11622

Keywords: Recurrent pregnancy loss, Taqman real-time PCR, Waddlia chondrophila

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.