No abstract

Summary Human papillomavirus DNA was detected in cervical specimens from 366 sexually active young women with cytomorphologically normal cervices using the polymerase chain reaction. In 93% (25/27) of initially infected women, the same viral type was not detected upon re-examination four menstrual cycles later. These results suggest that the majority of HPV infections in young women are transient.Keywords: human papillomavirus; polymerase chain reaction During the last decade considerable evidence has accumulated indicating a central role for human papillomaviruses (HPVs) in the aetiology of cervical cancer (zur Hausen, 1991;Schiffman, 1992). Current knowledge regarding the natural history of HPV infection is limited. Cross-sectional studies have shown that cervical HPV infection is common among women with normal cervicovaginal cytology; point prevalence measurements in such women were seen to peak in sexually active teenagers and women in their early twenties and then to decrease substantially with increasing age (Ley et al., 1991;Melkert et al., 1993;Morrison et al., 1991). This early peak in the prevalence of detectable HPV DNA in cytologically normal women contrasts with the peak prevalence of high-grade cervical intraepithelial neoplasia (CIN) occurring 5-10 years later and the plateau of invasive cancer observed 20 years subsequently. On the basis of these crosssectional data it was speculated that infection with HPV frequently occurs within a few years of the onset of sexual activity, running a transient self-limiting course in the majority of infected women and a chronic or recurrent course in a minority who may ultimately develop cervical neoplasia (Schiffman, 1992;Morrison, 1994 Specimen collection Combined cytobrush-spatula sampling of the cervix was performed on two occasions, before (cycle 0) and four menstrual cycles after (cycle 4), device insertion. On each occasion a Papanicolaou smear was sent for standard cytological assessment and HPV detection was performed using the polymerase chain reaction. Smears were assessed by one of the authors (MEB), without knowledge of the results of HPV PCR.Detection of HP V DNA Detection of HPV DNA was performed directly on cells suspended in Kryofix (Merck, product no. 5201) using a combination of general primer-mediated and type-specific PCR (GP/TS-PCR) (Walboomers et al., 1992). This combination of PCR techniques allows the rapid, sensitive and reliable detection of a broad spectrum of HPV genotypes in cervical cell suspensions . GP/TS-PCR facilitates the testing of large groups of smears and is considered the technique of choice for epidemiological studies (Schiffman, 1992).Initially, PCR was performed using general HPV primers GP 5/6 to determine the overall presence of HPV. This screening detects presently unsequenced genital HPV types, at the subpicogram level, in addition to the sequenced genital

Vaginal lactobacilli assessed by PCR-based microarray and PCR-based genotyping of HPV in South African women at risk for HIV and BV. Vaginal lactobacilli can be defined by microarray techniques in fixed cervical samples of South African women. Cervical brush samples suspended in the coagulant fixative BoonFix of one hundred women attending a health centre for HIV testing in South Africa were available for this study. In the Ndlovu Medical Centre in Elandsdoorn, South Africa, identification of 18 hr-HPV genotypes was done using the INNO-LiPA method. An inventory of lactobacilli organisms was performed using microarray technology. On the basis of the Lactobacillus and Lactobacillus biofilm scoring, the cases were identified as Leiden bacterial vaginosis (BV) negative (BV-; n = 41), Leiden BV intermediate (BV±; n = 25), and Leiden BV positive (BV+; n = 34). Fifty-one women were HIV positive and 49 HIV negative. Out of the 51 HIV positive women, 35 were HPV infected. These 51 HIV positive women were frequently infected with HPV16 and HPV18. In addition, HPV35, HPV52, HPV33, and HPV66 were often detected in these samples. Lactobacillus salivarius and Lactobacillus iners were the most prevalent lactobacilli as established by the microarray technique. In women with HPV infection, the prevalence of Lactobacillus crispatus was significantly reduced. In both HIV and HPV infection, a similar (but not identical) shift in the composition of the lactobacillus flora was observed. We conclude that there is a shift in the composition of vaginal lactobacilli in HIV-infected women. Because of the prominence of HPV35, HPV52, HPV33, and HPV66, vaccination for exclusively HPV16 and HPV18 might be insufficient in South African HIV+ women.

GPs often diagnose "vulvovaginal candidiasis" in their practices, but often do not perform the laboratory examinations required to confirm their putative diagnosis. This may lead to wrong diagnoses and maltreatment with antimycotics, without cure of the patients' vaginal complaints.

BackgroundExternal quality assurance (EQA) programmes, which are routinely used in laboratories, have not been widely implemented for point-of- care tests (POCTs). A study was performed in ten health centres in Tanzania, to implement the use of dried blood spots (DBS) as an EQA method for HIV and syphilis (POCTs).MethodDBS samples were collected for retesting at a reference laboratory and the results compared to the POCT results obtained at the clinic. In total, 2341 DBS samples were collected from 10 rural health facilities over a period of nine months, of which 92.5% were correctly collected and spotted.ResultsThe EQA method was easily implemented by healthcare workers under routine conditions in Northern Tanzania. For HIV, 967 out of 972 samples (99.5%) were concordant between DBS and POCT results. For syphilis, the sensitivity of syphilis tests varied between clinics with a median of 96% (25th and 75th quartile; 95-98%). The specificity of syphilis POCT was consistent compared to laboratory based test using DBS, with a median of 96% (25th and 75th quartiles; 95-98%).ConclusionOverall, the quality of testing varied at clinics and EQA results can be used to identify clinics where healthcare workers require remedial training, suggesting the necessity for stringent quality assurance programmes for POC testing. As Tanzania embarks on scaling up HIV and syphilis testing, DBS can be a useful and robust tool to monitor the quality of testing performed by healthcare workers and trigger corrective action to ensure accuracy of test results.