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...

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Objective: Some HIV+ patients, virally suppressed on ART, show occasional ‘blips’ of detectable HIV-1 plasma RNA. We used a new highly sensitive assay of cell-associated HIV-1 RNA to measure transcriptional activity in PBMCs and production of infectious virus from the viral reservoir, in patients with and without ‘blips’. Design/methods: RNA and DNA extracted from cells in 6 ml of peripheral blood, from suppressed patients with one to two ‘blip’ episodes over the past 2 years of ART ( n = 55), or no ‘blips’ ( n = 52), were assayed for HIV-1 RNA transcripts and proviral DNA targeting the highly conserved ‘R’ region of the LTR. Follow-up samples were also collected. Purified CD4 + T cells were cultured with anti-CD3/CD28/CD2 T-cell activator to amplify transcription and measure replication competent virus. Results: HIV-1 RNA transcripts ranged from 1.3 to 5415 copies/10 6 white blood cells. ‘Blip’ patients had significantly higher levels vs. without blips (median 192 vs. 49; P = 0.0007), which correlated with: higher levels of inducible transcripts after activation in vitro , sustained higher HIV-1 transcription levels in follow-up samples along with increasing HIV-1 DNA in some, and production of replication-competent HIV-1. Conclusion: Viral ‘blips’ are significant reflecting higher transcriptional activity from the reservoir and contribute to the reservoir over time. This sensitive assay can be used in monitoring the size and activity of the HIV-1 reservoir and will be useful in HIV-1 cure strategies.

Objectives Trichomonas vaginalis (TV) has a low profile in urban sexually transmitted infection (STI) clinics in many developed countries. The objective of this study was to determine the true prevalence of TV in an Australian urban sexual health setting using sensitive molecular diagnostic techniques. Methods A cross-sectional study investigating the aetiology of cervicitis in women attending two urban sexual health clinics in Sydney, Australia, enrolled 356 consecutive eligible women from 2006 to 2008. The diagnostic yield from the standard clinical practice of discretionary high vaginal wet preparation microscopy in women with suspicious vaginal discharge was compared with universal use of nested PCR for TV of cervical samples. Results TV was detected by PCR in 17/356 women (4.8%, 95% CI 2.8 to 7.5%), whereas only four cases (1.1%, 95% CI 0.3 to 2.8%) were detected by discretionary wet preparation microscopy. Eleven of the 17 women (p¼0.003) were of culturally and linguistically diverse background. Additionally, cervicitis was found to be significantly associated with TV, RR 1.66 (1.14 to 2.42), p¼0.034. Conclusions Traditional TV-detection methods underestimate TV prevalence in urban Australia. The TV prevalence of 4.8% by PCR testing in this study exceeds previously reported urban Australian TV rates of <1%. An increase in trichomoniasis-associated adverse reproductive outcomes and enhanced HIV transmission poses a salient public health threat. Accordingly, TV warrants a higher profile in urban STI clinic settings in developed countries, and we suggest that priority be given to development of standardised molecular TV detection techniques and that these become part of routine STI testing.