To Establish A Taqman MGB Real Time PCR For Detection Of Mycoplasma Genitalium Among High Risk Populations In China

Mycoplasma genitalium (Mg) was first isolated from the urethras of two men with nongonococcal urethritis (NGU) in1980, and it has been widely recognized as a sexually transmitted pathogen and one of the common causes of NGU. Mg is associated with cervicitis, endometritis, and pelvic inflammatory diseases (PID) in women, if left untreated, Mg infection can lead to tubal factor infertility.In China, NGU has been resurgent since1980s and used to be one of sexually transmitted infections (STIs) for case-reporting before2008. Since then NGU has been replaced by Chlamydia trachomatis (CT) for national sentinel surveillance programme. A few studies on Mg were conducted in China, showing a prevalence rate of20.6%to25%among NGU patients, and44.19%among non-chlamydial NGU patients. These findings indicate that Mg infection is prevalent in China.Even though the overall HIV prevalence in China is low, HIV infection through heterosexual transmission is increasing. Female sex workers, male STD clinic patients and men who have sex with men are high risk populations and potential bridge populations for STD and HIV infection and transmission. A significant association between Mg and HIV infection has been observed in many cross-sectional studies and is well-reviewed. In a systematic review and meta-analysis of19studies, Mg infection was associated with a2-fold increased risk of HIV infection. Although Mg prevalence in high-risk populations such as female sex workers and men who have sex with men is gaining increasing attention worldwide, only a few studies estimated Mg prevalence among small Chinese female sex workers, male STD clinic patients and men who have sex with men. This report is the first to determine the prevalence and risk factors for Mg among relative large sample size of these three high risk populations in southwest China.Sectionl:To establish a Taqman MGB real time PCR for detection of Mycoplasma genitaliumTo development a previously published and validated method of real-time PCR targeting the mgpA gene in our laboratory by using the standard Mg G37strain (ATCC33530) to build a standard curve, two important values:limit of detection as well as limit of quantification were estimated, the results indicated that the Taqman MGB Real time PCR had good linear range (1×101copies/μl---1×106copies/μL R2=0.993), great repeatability, and very sensitive with limit of detection was2copies/μl, limit of quantification was10copies/μl. We concluded that the Taqman MGB Real time PCR was a rapid and sensitive method which was available not only for detection but also for quantification, it may play an important role in detecting and monitoring Mg prevalence in high risk populations.Section2:High prevalence and risk factors for Mycoplasma genitalium among female sex workers and male STD clinic patients in Guangxi, China.The study populations included females who were older than18years and reported having provided commercial sex in the past year and male adults who attended in STD clinics. From July to September2009,810venue-based FSWs were recruited in two cities, Wuzhou and Hezhou,423male STD clinic patients were recruited in Hezhou of Guangxi Autonomous Region. After informed content was obtained, information regarding socio-demographic and behavioral characteristics was collected by a questionnaire-based interview by outreach workers. Cervical swab specimens and urine specimens were collected from female sex workers and male STD clinic patients respectively. Mg was detected by using a real time polymerase chain reaction assay targeting the mgpA gene. The overall prevalence of Mg infection among810FSWs was13.2%(95%CI=10.87%-15.52%). Mg infection rates among high-tier FSWs, middle-tier FSWs and low-tier FSWs were15.7%(95%CI=13.12%-18.2%),10.8%(95%CI=8.67%-12.93%), and14.6%(95%CI=12.17%-17.03%), respectively. Mg infection was only independently associated with less education (AOR=2.36,95% CI1.15-4.87), being single (AOR=2.27,95% CI=1.42-3.62), migrant backgrounds (AOR=2.03,95% CI=1.29-3.20), and absence of STIs symptoms in the previous year (AOR=1.66,95%CI=1.09-2.52). For MSP, Mg infection rate was28%(95%CI=25.50%-30.49%). Mg infection related to living alone (AOR=3.84,95%CI=1.58-9.33), ever had sex with FSWs within3months (AOR=2.27,95%CI=1.41-3.67), and absence of STIs symptoms in the previous year (AOR=2.21,95%CI=1.35-3.62) among MSP. In this study, the overall prevalence of Mg infection was found to be substantial among female sex workers as well as male STD clinic patients in study areas. These findings highlight the needs for increased attention towards prevention and intervention of Mg infection among these two high risk populations. Routine screening, timely diagnosis, and treatment of Mg infection should be considered in comprehensive HIV/STIs control program in China. Section3:The prevalence of urethral Mycoplasma genitalium and its associated factors among men who have sex with men in Guangzhou, ChinaMen who have sex with men from different venues including:gar bars, sauna, night clubs and parks were recruited by snowball method in Guangzhou, China, during Jan to May2010. Socio-demographic and behavioral characteristics information was collected by a questionnaire-based interview by outreach workers. Urine specimens were collected. Mg was detected by a real time PCR targeting the MgPa gene. Among328MSM, majority of study subjects (97.6%) were self-identified as gay, however, among them41.8%also had sex with women,16.8%of study subjects reported had sex with women within the previous6months,27.3%used condom during the last intercourse with women. Over half (58.8%) of study subjects only had sex with men, majority (87.8%) of study subjects had sex with men within the previous6months, and63.2%used condom during the last anal sex. Out of all MSM,79.0%solicited sex partners by internet.323urine specimens were collected, Mg infection rate was4.3%(95%CI=3.1%-5.4%). These findings indicate that Mg is relatively low prevalent among MSM, which suggests that Mg infection mainly by heterosexual transmission other than homosexual transmission, however MSM as source of Mg infection should not be neglected.