Multiplex PCR Testing Detection of Higher-than-Expected Rates of Cervical Mycoplasma , Ureaplasma , and Trichomonas and Viral Agent Infections in Sexually Active Australian Women

Abstract

Knowing the prevalence of potential etiologic agents of nongonococcal and nonchlamydial cervicitis is important for improving the efficacy of empirical treatments for this commonly encountered condition. We describe four multiplex PCRs (mPCRs), designated VDL05, VDL06, VDL07, and VDL09, which facilitate the detection of a wide range of agents either known to be or putatively associated with cervicitis, including cytomegalovirus (CMV), enterovirus (EV), Epstein-Barr virus (EBV), varicella-zoster virus (VZV), herpes simplex virus type 1 (HSV-1), and herpes simplex virus type 2 (HSV-2) (VDL05); Ureaplasma parvum, Ureaplasma urealyticum, Mycoplasma genitalium, and Mycoplasma hominis ( Cervicitis, an acute or chronic inflammation of the uterine cervix, is generally viewed as a consequence of infection with sexually transmissible agents. Neisseria gonorrhoeae and Chlamydia trachomatis are the most commonly reported pathogens, possibly because they are most frequently screened for. However, the etiology of most cases is undetermined and could be multifactorial in nature (11,34,35,40). Studies undertaken in other epidemiologic settings indicate significant differences in the prevalences of other cervical infectious agents (1,41,44,45,58). An underappreciation of the prevalences of and roles played by these nongonococcal and nonchlamydial agents potentially jeopardizes the effectiveness of empirical treatments for cervicitis. Unresolved cervicitis can result in ascending infection, endometritis, pelvic inflammatory disease, and salpingitis (11,23,46). Furthermore, cervicitis may enhance human immunodeficiency virus susceptibility by the disruption of mucosa, allowing increased viral replication within recruited inflammatory cells (30). The development of molecular methods, such as PCR and DNA hybridization, has allowed the detection of a range of agents whose etiologic roles in genital infections need to be further investigated, including the viruses cytomegalovirus (CMV), herpes simplex virus type 1 (HSV-1) and HSV-2 (4, 43), adenovirus (6,10,50), and the Mollicutes Ureaplasma parvum, Ureaplasma urealyticum, Mycoplasma hominis, and Mycoplasma genitalium (1,28,59). There have also been reports of genital infections caused by Epstein-Barr virus (EBV) (4, 55), varicella-zoster virus (VZV) (27), and enterovirus (EV) (24). We report here the use of four multiplex PCR (mPCR) assays, designated VDL05, VDL06, VDL07, and VDL09, based on a conventional platform, for the detection of 19 microorganisms in cervical swabs, including Treponema pallidum and C. trachomatis, Trichomonas vaginalis, group B streptococci, and five adenovirus species, in addition to those mentioned above. The assays were developed using cervical swabs from different women taken on one or more occasions during different visits to a sexual-health clinic. MATERIALS AND METHODSPatients. Cervical swabs (n ϭ 233) were taken from 175 women consecutively attending a sexual-health clinic in Sydney, Australia (between one and three visits), during 2006 and 2007 who we...

MMP-9/RECK Imbalance: A Mechanism Associated with High-Grade Cervical Lesions and Genital Infection by Human Papillomavirus and Chlamydia trachomatis

“…Positive controls for all genital infections were derived from positive clinical samples diagnosed by reference methods, including culture and/or single PCR (sPCR; ref. 22). Human b-globin-specific primers GH20/PC04 were used as internal controls for amplification and DNA integrity.…”

Simultaneous Detection of Seven Sexually Transmitted Agents in Human Immunodeficiency Virus–Infected Brazilian Women by Multiplex Polymerase Chain Reaction

“…The single-target PCR is generally more sensitive than the multiplex PCR and the possibility of cross-reactions, which can occur during M-PCR, are avoided. 7 All clinical samples were also tested by using specific primers GH20/ PC04 for the human β-globin gene as an internal control of amplification and DNA integrity under the same conditions of M-PCR or single-target PCR. Statistical analysis was performed by using STATISTICA 8.0 software (StatSoft Inc., Tulsa, OK), and all variables were expressed as absolute and relative frequencies.…”

Answer to August 2011 Photo Quiz

“…For this reason, it is important for the laboratory staff to communicate their findings to the patient's physician in order to prompt further workup, including HIV testing, if the patient has concurrent risk factors (1). Another setting in which CMV cervicitis may be clinically significant is during pregnancy (2,3,5). This is due to the virus's association with early spontaneous abortion as well as fetal infection resulting in congenital chorioretinitis, calcification of the cerebrum, and microcephaly.…”

“…These inclusions can help distinguish CMV from HSV infection. Although the CMV cytopathic effect is a rare finding on cervical cytology, the actual carriage rate determined using molecular diagnostics can range from 14% to 29%, indicating that it may be routinely overlooked (4,5). Patients with CMV cervicitis are typically asymptomatic and do not require treatment.…”

“…This is due to the virus's association with early spontaneous abortion as well as fetal infection resulting in congenital chorioretinitis, calcification of the cerebrum, and microcephaly. Although current CDC guidelines do not recommend routine serologic screening for all pregnant women, studies suggest screening of women who are at high risk for primary infection, including health care and childcare workers (2,5).…”