Development of a Taqman MGB real time fluorescence-based PCR assay for the detection of Mycoplasma genitalium / 中华皮肤科杂志

ABSTRACT

ObjectiveTo establish a rapid,sensitive and accurate method to detect Mycoplasma genitalium,and to evaluate the prevalence of M.genitalium among unlicensed prostitutes from Hezhou city in Guangxi Zhuang Autonomous Region.MethodsA pair of primers and Taqman MGB probe were designed and synthesized for the Pa gene of M.genitalium.Standard samples were prepared with the M.genitalium type strain G37.The established Taqman MGB real time fluorescence-based PCR assay was used to detect M.genitalium in the standard samples and cervical swab specimens collected from unlicensed prostitutes in Hezhou city of Guangxi Zhuang Autonomous Region.ResultsThe established Taqman MGB real time PCR exhibited a wide linear range( 1 × 10 copies/μl to 1 × 106 copies/μl,R2 =0.993),good repeatability(intra-assay variation;0.7％,inter-assay variation1.09％) and hign sensitivity with the limit of detection being 10 copies/μl and limit of quantification being 50 copies/μl.As the assay showed,12.1％ of the 404 cervical swab samples were positive for M.genitalium.ConculsionThe Taqman MGB real time fluorescence-based PCR is a rapid and sensitive method for the quantitative and qualitative detection of M.genitalium.