Prevalence and risk factors of sexually transmitted infections and cervical neoplasia in women from a rural area of southern mozambique

There is limited information on the prevalence of sexually transmitted infections and the prevalence of cervical neoplasia in rural sub-Saharan Africa. This study describes the prevalence and the etiology of STIs and the prevalence of cervical neoplasia among women in southern Mozambique. An age-stratified cross-sectional study was performed where 262 women aged 14 to 61 years were recruited at the antenatal clinic (59%), the family-planning clinic (7%), and from the community (34%). At least one active STI was diagnosed in 79% of women. Trichomonas vaginalis was present in 31% of all study participants. The prevalence of Neisseria gonorrhea and Chlamydia trachomatis were 14% and 8%, respectively, and Syphilis was diagnosed in 12% of women. HPV DNA was detected in 40% of women and cervical neoplasia was diagnosed in 12% of all women. Risk factors associated with the presence of some of the STIs were being divorced or widowed, having more than one sexual partner and having the partner living in another area. A higher prevalence was observed in the reproductive age group and some of the STIs were more frequently diagnosed in pregnant women. STI control programs are a priority to reduce the STIs burden, including HIV and cervical neoplasia.

The Role of Human Papillomavirus Genotyping in Cervical Cancer Screening: A Large-Scale Evaluation of the cobas HPV Test.

BACKGROUND: The cobas HPV Test ("cobas"; Roche Molecular Systems) detects HPV16 and HPV18 individually, and a pool of 12 other high-risk (HR) HPV types. The test is approved for (i) atypical squamous cells of undetermined significance (ASC-US) triage to determine need for colposcopy, (ii) combined screening with cytology ("cotesting"), and (iii) primary HPV screening. METHODS: To assess the possible value of HPV16/18 typing, >17,000 specimens from a longitudinal cohort study of initially HPV-positive women (HC2, Qiagen) were retested with cobas. To study accuracy, cobas genotyping results were compared with those of an established method, the Linear Array HPV Genotyping Test (LA, Roche Molecular Systems). Clinical value of the typing strategy was evaluated by linking the cobas results (supplemented by other available typing results) to 3-year cumulative risks of CIN3+. RESULTS: Grouped hierarchically (HPV16, else HPV18, else other HR types, else negative), the κ statistic for agreement between cobas and LA was 0.86 [95% confidence interval (CI), 0.86-0.87]. In all three scenarios, HPV16-positive women were at much higher 3-year risk of CIN3+ than HPV16-negative women: women ages 21 and older with ASC-US (14.5%; 95% CI, 13.5%-15.5% vs. 3.5%; 95% CI, 3.3-3.6); women ages 30 years and older that were HPV-positive cytology-negative (10.3%; 95% CI, 9.6-11.1 vs. 2.3%; 95% CI, 2.2-2.4); and all women 25 years and older that were HPV-positive (18.5%; 95% CI, 17.8-19.2 vs. 4.3%; 95% CI, 4.2-4.4). CONCLUSION: The cobas and LA results show excellent agreement. The data support HPV16 typing. IMPACT: HPV16 typing is useful in the management of HPV-positive/cytology-negative women in cotesting, of all HPV-positive women in primary HPV testing, and perhaps in the management of HPV-positive women with ASC-US. Cancer Epidemiol Biomarkers Prev; 24(9); 1304-10.

Prevalence and risk factors of sexually transmitted infections and cervical neoplasia in women from a rural area of southern Mozambique.

There is limited information on the prevalence of sexually transmitted infections and the prevalence of cervical neoplasia in rural sub-Saharan Africa. This study describes the prevalence and the etiology of STIs and the prevalence of cervical neoplasia among women in southern Mozambique. An age-stratified cross-sectional study was performed where 262 women aged 14 to 61 years were recruited at the antenatal clinic (59%), the family-planning clinic (7%), and from the community (34%). At least one active STI was diagnosed in 79% of women. Trichomonas vaginalis was present in 31% of all study participants. The prevalence of Neisseria gonorrhea and Chlamydia trachomatis were 14% and 8%, respectively, and Syphilis was diagnosed in 12% of women. HPV DNA was detected in 40% of women and cervical neoplasia was diagnosed in 12% of all women. Risk factors associated with the presence of some of the STIs were being divorced or widowed, having more than one sexual partner and having the partner living in another area. A higher prevalence was observed in the reproductive age group and some of the STIs were more frequently diagnosed in pregnant women. STI control programs are a priority to reduce the STIs burden, including HIV and cervical neoplasia.

Genotyping of human papillomavirus DNA in anal biopsies and anal swabs collected from HIV-seropositive men with anal dysplasia.

BACKGROUND: Human papillomavirus (HPV) causes anal intraepithelial neoplasia (AIN) in HIV-seropositive men. The detection of HPV genotypes in anal biopsies and swabs was compared. METHODS: HPV DNA was detected in anal swabs and biopsies obtained concurrently from 154 HIV-seropositive men [31 without AIN, 60 low-grade AIN (AIN-1), 62 high-grade AIN (AIN-2,3), and 1 indeterminate AIN] under or eligible to highly active antiretroviral therapy. RESULTS: HPV DNA was detected in 24.2% of normal biopsies compared with 93.5% with AIN-2,3 (P < 0.001) and 88.3% with AIN-1 (P < 0.001). The proportion of biopsies containing multiple genotypes was greater in AIN-1 (n = 21, 35.0%; P = 0.002) and AIN-2,3 (n = 38, 58%; P < 0.001) than in normal biopsies (n = 2, 6.5%). The most frequent genotypes in order of frequency were in AIN-2,3 biopsies HPV-16, 18, 58, and 45 and were in AIN-1 biopsies HPV-6, 11, 16, and 39. Controlling for age, CD4 count, and smoking, the presence of high-risk HPV DNA in biopsies [odds ratio (OR) = 50.8, 95% confidence interval (CI): 13.0 to 199.5] but not in swabs (OR = 2.0, 95% CI: 0.6 to 7.0) was associated with AIN-2,3. CONCLUSIONS: AIN-2,3 was associated with high-risk HPV infection detected in biopsies but not in swabs in men under or starting highly active antiretroviral therapy, possibly due to the presence of HPV foci outside of the neoplastic lesion.

Mouthwash as a low-cost and safe specimen transport medium for human papillomavirus DNA testing of cervicovaginal specimens.

The usefulness of mouthwash as a transport medium for cervical specimens for carcinogenic human papillomavirus (HPV) DNA testing has not been evaluated. Two cervical specimens were collected from each of 34 patients, with one placed in mouthwash (Scope, Proctor and Gamble, Inc.) and the other in a liquid cytology medium commonly used for HPV DNA testing in alternating order. Paired specimens were tested by a PCR assay for carcinogenic HPV and a PCR HPV genotyping assay for 37 HPV types at 0, 3, and 6 weeks after collection; the results of the HPV genotyping assay were categorized into HPV risk groups according to cancer risk (HPV-16 > HPV-18 > other carcinogenic HPV types > noncarcinogenic HPV types > negative). After 4 months of storage, specimens were tested using a second, non-PCR test for carcinogenic HPV. We observed a >or=94% total agreement and kappa values of >or=0.88 between media at each time point for PCR-detected carcinogenic HPV. We observed a >or=74% total agreement, >or=0.62 unweighted kappa, and >or=0.75 linearly weighted kappa between media at each time point for PCR-detected HPV cancer risk category. Finally, we observed an 88% total agreement and kappa of 0.77 between media for carcinogenic HPV detection using a second test after 4 months of storage. We suggest that mouthwash might be used as a low-cost, safe, nonflammable storage and transport medium for cervical specimens for HPV DNA testing in cervical cancer screening programs.

Enhanced detection and typing of human papillomavirus (HPV) DNA in anogenital samples with PGMY primers and the Linear array HPV genotyping test.

The Roche PGMY primer-based research prototype line blot assay (PGMY-LB) is a convenient tool in epidemiological studies for the detection and typing of human papillomavirus (HPV) DNA. This assay has been optimized and is being commercialized as the Linear Array HPV genotyping test (LA-HPV). We assessed the agreement between LA-HPV and PGMY-LB for detection and typing of 37 HPV genotypes in 528 anogenital samples (236 anal, 146 physician-collected cervical, and 146 self-collected cervicovaginal swabs) obtained from human immunodeficiency virus-seropositive individuals (236 men and 146 women). HPV DNA was detected in 433 (82.0%) and 458 (86.7%) samples with PGMY-LB and LA-HPV (P = 0.047), respectively, for an excellent agreement of 93.8% (kappa = 0.76). Of the 17,094 HPV typing results, 16,562 (1,743 positive and 14,819 negative results) were concordant between tests (agreement = 96.9%; kappa = 0.76). The mean agreement between tests for each type was 96.4% +/- 2.4% (95% confidence interval [CI], 95.6% to 97.2%; range, 86% to 100%), for an excellent mean kappa value of 0.85 +/- 0.10 (95% CI, 0.82 to 0.87). However, detection rates for most HPV types were greater with LA-HPV. The mean number of types per sample detected by LA-HPV (4.2 +/- 3.4; 95% CI, 3.9 to 4.5; median, 3.0) was greater than that for PGMY-LB (3.4 +/- 3.0; 95% CI, 3.1 to 3.6; median, 2.0) (P < 0.001). The number of types detected in excess by LA-HPV in anal samples correlated with the number of types per sample (r = 0.49 +/- 0.06; P = 0.001) but not with patient age (r = 0.03 +/- 0.06; P = 0.57), CD4 cell counts (r = 0.06 +/- 0.06; P = 0.13), or the grade of anal disease (r = -0.11 +/- 0.06; P = 0.07). LA-HPV compared favorably with PGMY-LB but yielded higher detection rates for newer and well-known HPV types.

Prevalence and determinants of human papillomavirus infection in men attending vasectomy clinics in Mexico.

Large studies of genital human papillomavirus (HPV) infection in men are few and mainly include high-risk groups. We interviewed 779 men who requested a vasectomy in 27 public clinics in 14 states of Mexico. Exfoliated cells were obtained from the scrotum, the shaft of the penis, the top of the penis including the coronal sulcus, the glans and the opening of the meatus. HPV testing was performed using biotinylated L1 consensus primers and reverse line blot. Unconditional logistic regression was used to estimate odds ratios (ORs) of being HPV-positive and corresponding 95% confidence intervals (CIs). The prevalence of any type of HPV was 8.7%. HPV positivity was highest among men below age 25 (13.6%), and lowest among men aged 40 years or older (6.0%). The most commonly found HPV types were, in decreasing order, HPV59, 51, 6, 16 and 58. Lifetime number of sexual partners was associated with HPV positivity (OR for > or = 4 vs. 1 partner = 3.7, 95% CI: 2.0-6.8), mainly on account of the strong association with number of occasional and sex-worker partners. Condom use with both regular (OR = 0.4, 95% CI: 0.1-1.0) and sex-worker (OR = 0.1, 95% CI: 0.0-0.3) partners and circumcision (OR = 0.2, 95% CI: 0.1-0.4) were inversely associated with HPV positivity. HPV prevalence in Mexican men was similar to the prevalence found in Mexican women of the same age groups. The association between HPV positivity and lifetime number of sexual partners in the present low-risk male population is one of the strongest ever reported in studies in men. Condom use and circumcision were associated with a strong reduction in HPV prevalence.

Evaluation of a modified reverse line blot assay for detection and typing of human papillomavirus.

Detection and typing of human papillomaviruses (HPVs) are requested by clinicians with growing frequency as part of the overall management of patients. A new method using consensus L1 amplification and a reverse line blot hybridization detection method has been described for broad spectrum HPV genotype identification. This method involves hybridization at 53 degrees C in a shaking water bath, which many laboratories might find cumbersome. We describe a modified hybridization step, using a dry-air incubator, that simplifies the detection protocol. Overall, comparable results were obtained by the modified, more easily performed protocol, indicating that laboratories might validate and use this modified method.

Evaluation of a convenient enzyme immunoassay to assess the quality of genital specimens submitted for the detection of human papillomavirus DNA by consensus PCR.

BACKGROUND: In the PGMY-line blot assay, a human beta-globin fragment is co-amplified with human papillomavirus (HPV) DNA, and both analytes are detected by hybridization with probes fixed on a strip in a linear array. The beta-globin DIG-MWP test also detects beta-globin amplicons, but in a microtiter plate-based enzyme immunoassay format. Although the PGMY-line blot assay detected 50 cells per test, the beta-globin DIG-MWP test generated a signal above the detection cut-off with five cells per test. OBJECTIVE: The performance of the beta-globin DIG-MWP assay to detect beta-globin DNA was assessed. STUDY DESIGN: The beta-globin DIG-MWP assay was compared to a standard beta-globin PCR and to the PGMY-line blot strips on 401 genital specimens. Overall, the three beta-globin assays were compared on 325 undiluted lysates, 14 diluted lysates and DNA extracted from 62 lysate samples. RESULTS: Concordance between the PGMY-line blot and the standard beta-globin assay reached 99.5% (399 of 401 results), for a kappa value of 0.95. Concordant results were also obtained between the beta-globin DIG-MWP assay and PGMY-line blot assay for 387 (96.5%) of 401 test results, for a kappa value of 0.57. Discordant results were due to the increased sensitivity of the DIG-MWP assay. Using a cut-off for positivity at 1.500 optical density (OD) units for beta-globin DIG-MWP, concordance improved to 100% (401 of 401 results, kappa at 1.00). CONCLUSION: The beta-globin DIG-MWP assay was adequate to screen for sample adequacy for HPV analysis in genital specimens.

International proficiency study of a consensus L1 PCR assay for the detection and typing of human papillomavirus DNA: evaluation of accuracy and intralaboratory and interlaboratory agreement.

The PGMY L1 consensus primer pair combined with the line blot assay allows the detection of 27 genital human papillomavirus (HPV) genotypes. We conducted an intralaboratory and interlaboratory agreement study to assess the accuracy and reproducibility of PCR for HPV DNA detection and typing using the PGMY primers and typing amplicons with the line blot (PGMY-LB) assay. A test panel of 109 samples consisting of 29 HPV-negative (10 buffer controls and 19 genital samples) and 80 HPV-positive samples (60 genital samples and 20 controls with small or large amounts of HPV DNA plasmids) were tested blindly in triplicate by three laboratories. Intralaboratory agreement ranged from 86 to 98% for HPV DNA detection. PGMY-LB assay results for samples with a low copy number of HPV DNA were less reproducible. The rate of intralaboratory agreement excluding negative results for HPV typing ranged from 78 to 96%. Interlaboratory reliability for HPV DNA positivity and HPV typing was very good, with levels of agreement of >95% and kappa values of >0.87. Again, low-copy-number samples were more prone to generating discrepant results. The accuracy varied from 91 to 100% for HPV DNA positivity and from 90 to 100% for HPV typing. HPV testing can thus be accomplished reliably with PCR by using a standardized written protocol and quality-controlled reagents. The use of validated HPV DNA detection and typing assays demonstrating excellent interlaboratory agreement will allow investigators to better compare results between epidemiological studies.

Human papillomavirus infection in men attending a sexually transmitted disease clinic.

Human papillomavirus (HPV) is the main etiologic agent of anogenital cancers, including cervical cancer, but little is known about the type-specific prevalence of HPV in men. Participants were men aged 18-70 years attending a sexually transmitted disease clinic. Penile skin swabs were assessed for HPV DNA using polymerase chain reaction with reverse line-blot genotyping. Of 436 swabs collected, 90.1% yielded sufficient DNA for HPV analysis. Men with inadequate swab samples were significantly more likely to be white and circumcised than men with adequate swab samples. The prevalence of HPV was 28.2%. Oncogenic HPV types were found in 12.0% of participants, nononcogenic types were found in 14.8% of participants, multiple types were found in 6.1% of participants, and unknown types were found in 5.9% of participants. The most prevalent subtypes were nononcogenic 6, 53, and 84. HPV positivity was not associated with age. These results indicate that HPV infection among men at high risk is common but that characteristics of male HPV infection may differ from those of female infection.