The accuracy and usefulness of laboratory-developed real-time PCR procedures using a Light Cycler instrument (Roche Diagnostics) for detecting and quantifying human immunodeficiency virus type 1 (HIV-1) RNA and DNA as well as herpes simplex virus type 1 (HSV-1)/HSV-2 DNA in cervicovaginal secretions from women coinfected with HIV and HSV were evaluated. For HIV-1, the use of the NEC152 and NEC131 primer set and the NEC-LTR probe in the long terminal repeat gene allowed us to detect accurately the majority of HIV-1 subtypes of group M circulating in sub-Saharan Africa, including subtypes A, B, C, D, and G as well as circulating recombinant forms 02 and 11. The detection threshold of real-time PCR for HIV in cervicovaginal lavage samples was 5 copies per assay for both RNA and DNA; the intra-and interassay coefficients of variation of C T values were 1.30% and 0.69% (HIV-1 RNA) and 1.84% and 0.67% (HIV-1 DNA), respectively. Real-time PCR for HSV using primers and probe targeting the HSV DNA polymerase gene allowed both detection and quantification of HSV DNA and also differentiation between HSV-1 and HSV-2 genotypes. The detection threshold of real-time PCR for HSV was 5 copies per assay; the intra-and interassay coefficients of variation of C T values were 0.96% and 1.49%, respectively. Both manual and automated silica-based procedures were appropriate for combined extraction of HIV and HSV genomes from female genital secretions. Taken together, these findings indicate that real-time PCR may be used as a unique nucleic acid amplification procedure to detect and quantify HIV and HSV genomes in cervicovaginal secretions and thus to assess at reduced costs the genital shedding of both viruses in women included in intervention studies.
BackgroundHigh-risk (HR) human papillomavirus (HPV) infection remains a great concern in relation to African men who have sex with men (MSM), especially those infected with HIV. The prevalence of HR-HPV and associated risk factors was estimated in a cross-sectional observational study covering MSM living in Bangui, Central African Republic.MethodsMSM receiving care at the Centre National de Référence des Infections Sexuellement Transmissibles et de la Thérapie Antirétrovirale, Bangui, were included. HIV serostatus and socio-demographic and behavioral characteristics were collected. HPV DNA was detected and genotyped on anal swabs using Anyplex™ II HPV28 test (Seegene, South Korea), and HSV DNA by in-house real-time PCR. Logistic regression analyses were used to determine risk factors associated with HPV outcomes.Results42 MSM (mean age, 23.2 years; range, 14–39) including 69.1% HIV-1-positive and 30.9% HIV-negative were prospectively enrolled. The prevalence of anal HPV was 69.1%, including 82.7% of HR-HPV which were multiple in 52.0%. The most prevalent genotypes were HPV-35, HPV-58, HPV-59 and HPV-31. While, HPV-16 and HPV-18 were present in a minority of samples. Multiple HR-HPV infection was more frequent in HIV-positive MSM (41.4%) with 2.7 genotypes per anal samples than in HIV-negative (7.7%) with 1.5 genotypes per anal samples. HPV types included in the prophylactic Gardasil-9® vaccine were detected in 68.9% of specimens and HPV-58 was the most frequently detected. MSM infected by HPV-16 and HPV-18 were all infected by HIV-1. Few anal swabs (11.9%) contained HSV-2 DNA without relationship with HPV detection. Condomless receptive anal intercourse was the main risk factor to being infected with any type of HPV and condomless insertive anal intercourse was significantly less associated with HPV contamination than receptive anal intercourse (Odd ratio = 0.02).ConclusionMSM in Bangui are at-risk of HIV and HR-HPV anal infections. The unusual distribution of HPV-35 as predominant HPV suggests possible geographic specificities in the molecular epidemiology of HR-HPV in sub-Saharan Africa. Scaling up prevention strategies against HPV infection and related cancers adapted for MSM in Africa should be prioritized. Innovative interventions should be conceived for the MSM population living in Bangui.
Background:Opportunities for HIV testing could be enhanced by offering HIV self-testing (HIVST) in populations that fear stigma and discrimination when accessing conventional HIV counselling and testing. Field experience with HIVST was poorly reported in French-speaking African countries.Objective:To investigate the usability of HIVST in Bangui, Central African Republic.Methods:The prototype self-test Exacto® Test HIV (Biosynex, Strasbourg, France) was used to assess the usability of HIVST in 300 adults living in Bangui, according to WHO technical recommendations. Simplified and easy-to-read leaflet was translated in French and Sango.Results:Preliminary survey in 3,484 adult volunteers including students, men who have sex with men and female sex workers living in Bangui showed that previous HIV testing in conventional centres for HIV counselling and testing was relatively infrequent and that acceptability of HIVST was elevated, although high heterogeneity could be observed between groups. The notice in French and Sango of Exacto® Test HIV were chosen in 242/300 (80.6%) and 58/300 (19.4%), respectively. It was correctly understood in 273/300 (91.0%). The majority (275/300; 91.6%) correctly performed the HIV self-test; however, 71/300 (23.0%) asked for oral assistance. Most of the participants (273/300; 91.0%) found that performing of the self-test was very easy or easy, and less than Most of participants (273/300; 91.0%) found that performing of the self-test was very easy or easy and less than 1.0% (2/300) found it difficult. Overall the result were correctly interpreted in 96.9% (3,782/3,900), the reading/interpretion errors concerned the positive (96/1,800;5.3%), invalid (17/600;2.8%) and negative (5/1,500; 0.3%) self-test. The Cohen's coefficient κwas 0.94. The main obstacle for HIVST was the educational level, with interpretation difficulties in poorly educated people.Conclusions:Our observations on profane adults living in Central African Republic, demonstrate: (i) the need to adapt the notice of instruction to African public, including educational pictograms as well as notice in vernacular language(s); (ii) the frequent difficulties in understanding the notice with frequent misinterpretation of test results; (iii) and the generally good usability of the HIV self-test despite these latter pitfalls. More research on exploring the best strategy (i.e. supervised versus unsupervised strategies) for different high- and low- risk populations in resource-constrained settings remains needed.
A total of 242 HIV-1-infected children were followed up at the Complexe Pédiatrique of Bangui, Central African Republic, including 165 receiving antiretroviral treatment in first- (n=150) or second-/third-line (n=15) regimens. They were prospectively included in a study, in 2009, to assess their virological status and prevalence of antiretroviral drug-resistance mutations in cases of virological failure, according to revised 2010 WHO criteria (e.g., HIV-1 RNA >3.7 log(10) copies/ml). Detectable plasma HIV-1 RNA was observed in 53% of children under first-line treatment, and virological failure was diagnosed in 40%, which was associated in 85% of cases with viruses harboring at least one drug-resistance mutation to nucleoside reverse transcriptase inhibitors (NRTI) or nonnucleoside reverse transcriptase inhibitors (NNRTI), and in 36% of cases with at least one major drug-resistance mutation to NRTI or NNRTI when excluding the M184V mutation. Overall, the proportion of children receiving a first-line regimen for a median of 18 months with virological failure associated with drug-resistance mutations, and thus eligible for a second-line treatment, was estimated at 34% of the whole cohort. In children under second-/third-line therapy, virological failure occurred in 47%, plus at least one major drug-resistance mutation to NRTI or NNRTI, though less commonly to protease inhibitors. Taken together, these findings argue in favor of the urgent need to improve distribution of pediatric antiretroviral drugs in the Central African Republic, to increase adherence by treated children, and to offer adequate HIV biological monitoring.
We report on field interpretation of HIV self-testing among female sex workers (FSWs) and non-FSWs living in Democratic Republic of the Congo. Two hundred and eight participants [76 (36.5%) FSWs; 132 (63.5%) non-FSWs] were enrolled in Kisangani and Bunia to evaluate their ability to read and interpret the results of a prototype HIV self-test (Exacto Test HIV, Biosynex, Strasbourg, France), according to WHO recommendations. Thirteen standardized tests (6 positive, 5 negative, 2 invalid) were proposed after successive random selection. Two thousand seven hundred and four standardized tests (1248 positive, 1040 negative, 416 invalid) were interpreted; 2435 (90.1%) were correctly interpreted, whereas 269 (9.9%) were misinterpreted. In FSWs and non-FSWs, the test results were similarly correctly interpreted in 87.4% (864/988) and 91.6% (1571/1716), respectively. In multivariate logistic regression analysis, only the variable “educational level” remained strongly associated with the interpretation of positive, negative, and invalid HIV self-test results, but not the variables “commercial sex work” and “language chosen for instructions for use.” Incorrect interpretation was significantly higher in participants with insufficient educational level than in those with sufficient education level for positive (13.1% vs 2.6%; adjusted OR: 4.5), negative (22.3% vs 2.6%; adjusted OR: 5.3), and invalid test results (23.8% v 6.4%; adjusted OR: 3.6). Incorrect interpretation of HIV self-test was as common in FSWs and non-FSWs. The lower was the educational level, the greater was the difficulty to interpret results correctly. These observations point that insufficient education level, rather than commercial sex work by itself, constitutes a key factor of incorrect interpretation of HIV self-test.
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