Genetic diversity of HPV35 in Chad and the Central African Republic, two landlocked countries of Central Africa: A cross-sectional study.

Human Papillomavirus (HPV)-35 accounts for up 10% of cervical cancers in Sub-Saharan Africa. We herein assessed the genetic diversity of HPV35 in HIV-negative women from Chad (identified as #CHAD) and HIV-infected men having sex with men (MSM) in the Central African Republic (CAR), identified as #CAR. Ten HPV35 DNA from self-collected genital secretions (n = 5) and anal margin samples (n = 5) obtained from women and MSM, respectively, were sequenced using the ABI PRISM® BigDye Sequencing technology. All but one HPV35 strains belonged to the A2 sublineage, and only #CAR5 belonged to A1. HPV35 from #CAR had higher L1 variability compared to #CHAD (mean number of mutations: 16 versus 6). L1 of #CAR5 showed a significant variability (2.29%), suggesting a possible intra-type divergence from HPV35H. Three (BC, DE, and EF) out of the 5 capsid loops domains remained totally conserved, while FG- and HI- loops of #CAR exhibited amino acid variations. #CAR5 also showed the highest LCR variability with a 16bp insertion at binding sites of the YY1. HPV35 from #CHAD exhibited the highest variability in E2 gene (P<0.05). E6 and E7 oncoproteins remained well conserved. There is a relative maintenance of a well conserved HPV35 A2 sublineage within heterosexual women in Chad and MSM with HIV in the Central African Republic.

Seroprevalence of anti-SARS-CoV-2 antibodies before and after implementation of anti-COVID-19 vaccination among hospital staff in Bangui, Central African Republic.

Healthcare workers (HCWs) are at high to very high risk for SARS-CoV-2 infection. The persistence of this pandemic worldwide has instigated the need for an investigation of the level of prevention through immunization and vaccination against SARS-CoV-2 among HCWs. The objective of our study was to evaluate any changes in anti-COVID-19 serological status before and after the vaccination campaign of health personnel in the Central African Republic. We carried out a repeated cross-sectional serological study on HCWs at the university hospital centers of Bangui. Blood samples were collected and tested for anti-SARS-CoV-2 IgM and IgG using the ELISA technique on blood samples. A total of 179 and 141 HCWs were included in the first and second surveys, respectively. Of these staff, 31.8% of HCWs were positive for anti-SARS-CoV-2 IgG in the first survey, whereas 95.7% were positive for anti-SARS-CoV-2 IgG in the second survey. However, the proportion of HCWs positive for SARS-CoV-2 IgM antibodies was low (9.7% in the first survey and 3.6% in the second survey). These findings showed a sharp increase in seroprevalence over a one-year period. This increase is primarily due to the synergistic effect of the infection and the implementation of vaccines against COVID-19. Further studies to assess the persistence of anti-SARS-CoV-2 antibodies are needed.

Effectiveness of Xpert MTB/RIF and the Line Probe Assay tests for the rapid detection of drug-resistant tuberculosis in the Central African Republic.

The Xpert MTB/RIF and Line Probe Assay (LPA) tests are more and more frequently used in mycobacteria testing laboratories for the rapid diagnosis of multi-drug resistance (MDR-TB). In this study, we demonstrate the effectiveness of these tests in the Central African Republic. Rifampicin resistance cases detected by the Xpert MTB/RIF during the year 2020 are also underwent first- and second-line LPA, and a first-line of drug susceptibility testing (DST) on solid medium and we compared these results. 101 rifampicin resistance cases based on the Xpert MTB/RIF were detected. Mean age was 34 years [16-81]. The 20-40 years age group represented 73.2% and the male-to-female sex ratio was 1.9:1. Patient profiles were dominated by treatment failure cases (40.6%) followed by relapsed cases (30.7%) and new cases (18.8%). These 101 rifampicin resistance were also detected with the first-line LPA and were confirmed by the DST. Similarly, the isoniazid results obtained with the first-line LPA, were confirmed by the DST, giving a concordance of 100% for these antibiotics. Rifampicin resistance were for the most part due to the absence of the WT8 sequence (56%) and the presence of the Mut3 mutation (53.4%). The majority of the isoniazid resistance (94.2%) were due to the Mut1 mutation in the katG gene and 4.2% of the cases involved both the katG gene and the inhA gene promoter with the Mut1 mutation. The second-line LPA test no resistance to second-line antibiotics. This study demonstrated the effectiveness of the Xpert MTB/RIF and the LPA tests for the rapid diagnosis of MDR-TB in the Central African Republic. However, due to their high cost, these tests have not been extensively deployed in the country. Public authorities and their TB-partners can help make these molecular tests more accessible to fight MDR-TB in the country.

Diagnostic performances of Exacto® Triplex rapid test for diagnosis of HIV/HCV/HBsAg: a multicenter, cross-sectional, field study in the Central African Republic.

Introduction: the Exacto® Triplex HIV/HCV/HBsAg (Biosynex, Strasbourg, France) consists in lateral flow, immunochromatographic rapid diagnostic test simultaneously detecting human immunodeficiency virus (HIV)-1 and HIV-2 and hepatitis C virus (HCV)- specific antibodies (IgG and IgM) and hepatitis B virus (HBV) surface antigen (HBsAg) in serum, plasma and whole blood. We herein evaluated its diagnostic performances in the Central African Republic (CAR). Methods: cross-sectional study was conducted on prospectively collected panel of 550 sera from adult inpatients living in Bangui, including 200 HIV-positive, 100 HBsAg-positive, 50 HCV-positive, 200 negatives to three viruses according to reference immuno-enzymatic serological tests including Murex HCV (Diasorin, Saluggia, Italy) for HCV, Murex HBsAg (Diasorin) for HBV, Genscreen ULTRA HIV Ag-Ab HIV-1/2 Version 2 (Bio-Rad, Marnes-la-Coquette, France) and Murex HIV 1.2.0 Ag/Ab Combination (Diasorin), the 2 tests associated in the parallel algorithm for the reference strategy for the diagnosis of HIV in CAR. Serum samples were tested blindly in duplicate. The findings are reported following the Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) guidelines. Results: the Exacto® Triplex showed 99.5% (95% CI; 98.5-100.0), 96.0% [90.6-100.0] and 99.0% [97.1-100.0] sensitivities for HIV, HCV and HBsAg, respectively. The specificity, positive and negative predictive values (PPV and NPV) were 100.0% for all three viruses. The Youden's J index and Cohen's κ coefficient were 0.99 for HIV and HBsAg. For HCV, Youden's J and Cohen's κ coefficient were 0.96 and 0.98, respectively. In the epidemiological context of the CAR, the PPV and NPV for all three viral infections were high (≥99.0% to 100%). Conclusion: taken together, our STROBE-compliant study demonstrates that the Exacto® Triplex HIV/HCV/HBsAg showed high sensitivity and specificity for HIV and HBsAg (≥99.0%), and relatively high sensitivity (96.0%) and high specificity (100%) for HCV. These analytical performances are within the limits required by the WHO (i.e. sensitivity ≥99.0% and specificity ≥98.0%) for HIV and HBV. The Exacto® Triplex HIV/HCV/HBsAg is user-friendly at low cost, and appears highly desirable for routine use in the CAR, and likely other Central African countries.

Analytical performances of Exacto® HIV self-test in the Central African Republic.

We herein evaluated the analytical performances of the CE-IVD capillary blood Exacto® HIV self-test (Biosynex, Strasbourg, France) in the Central African Republic (CAR). A cross-sectional study was conducted on a representative national panel of 200 sera positive for HIV and 200 negative for HIV, randomly selected thorough the CAR for HIV seroprevalence surveillance survey, according to reference test. The Exacto® HIV self-test showed 99.5% (95% CI: 98.2-99.9) sensitivity and 100.0% (95% CI: 99.0-100.0) specificity. The Youden´s J index and Cohen´s Kappa coefficient were 0.995. At HIV-1 seroprevalence of 3.5% in the general adult population of the CAR, the positive and negative predictive values were 100% (95% CI: 99.0-100) and 99.9% (95% CI: 98.9-100), respectively. The results are within the limits required by the WHO (i.e. sensitivity ≥ 99.0% and specificity ≥ 98.0%), making Exacto® HIV self-test suitable for routine use in the CAR.

Prévalence de l'hépatite virale B chronique à Bangui et Bimbo en République Centrafricaine: cas des donneurs bénévoles réguliers non éligibles pour les dons de sang / Prevalence of chronic viral hepatitis B in Bangui and Bimbo in the Central African Republic: cases of regular voluntary donors not eligible for blood donations

Contexte & objectif. L'hépatite virale B (HVB) par son évolution peut déboucher vers la guérison ou vers une forme chronique qui est très peu documentée chez les donneurs bénévoles de sang (DBS). L'objectif de la présente étude était de déterminer la prévalence de l'HVB chronique. Méthodes. L'étude réalisée au Centre National de Transfusion Sanguine de Bangui était rétrospective et portait sur les données de contrôle sérologique des DBS de Bimbo et de Bangui, capitale de la République centrafricaine (RCA). L'échantillonnage exhaustif concerne la période de juillet 2013 à décembre 2019. Le DBS est porteur d'une infection chronique si l'antigène de surface de l'HVB persiste pendant plus de six mois. Le test de Chi carré de Pearson au seuil de 5 % et l'odd ratio (OR) ont été utilisés comme test d'association. Résultats. Les données de contrôle sérologique de 702 DBS âgés de 18 à 62 ans ont été analysées. Le sexe masculin était prépondérant (n=598). La prévalence de l'HVB chronique était de 70,5 %. Cette prévalence était plus élevée chez les DBS de 25 à 34 ans (30,4 %), les hommes (58,8 %) et les DBS qui résident à Bangui (61,2 %). La forme chronique était significativement associée au jeune âge (18 à 44 ans) et au sexe masculin (p< 0,05). La coïnfection par le VIH et l'hépatite C était retrouvée dans 5,5 % des cas (39/702). Conclusion. La prévalence de l'HVB chronique est très élevée chez les DBS de Bangui et Bimbo. La forme chronique était significativement associée à l'âge et au sexe. La gratuité de la charge virale et du traitement antiviral sont des perspectives à mettre en œuvre

Context and objective. Viral hepatitis B (VHB) by its evolution can lead to recovery or to a chronic form. This chronic form, a source of new contaminations, is not documented among voluntary blood donors (VBD) in Bangui and Bimbo. The present study aimed to determine the prevalence of chronic VHB among VBD in the two cities in the Central African Republic (CAR). Methods. The study carried out at the Bangui National Blood Transfusion Center was retrospective and focused on serological control data from VBD from Bangui, capital of CAR and Bimbo. The exhaustive sampling is from July 2013 to December 2019. A VBD carries a chronic infection if the VHB surface antigen persists for more than six months. Results. Serological control data from 702 VBD aged 18 to 62 years were analyzed. Male sex predominated at inclusion (n = 598). The prevalence of chronic VHB was 70.5 %. This prevalence was higher among young VBD aged 25 to 34 years (30.4 %), the male gender (58.4 %) and VBD residing in Bangui (61.2 %). The chronic form was significantly associated with young age (18 to 44 years) and male sex (p< 5%). Co-infection with HIV and hepatitis C was found in 5.5 % of cases (39/702). Conclusion. The prevalence of chronic VHB is very high in VBD from Bangui and Bimbo. The chronic form was significantly associated with age and sex. Free viral load and antiviral treatment are prospects to be implemented.

Séroprévalence de la toxoplasmose chez les femmes en consultation prénatale à l'Hôpital du District de Bossembelé en République Centrafricaine en 2020 / Seroprevalence of toxoplasmosis in women in antenatal clinic at the Bossembelé District Hospital in the Central African Republic in 2020

Contexte et objectif. La toxoplasmose est une anthropozoonose ubiquitaire qui occupe une large place en médecine humaine et vétérinaire. Mais les données y relatives chez la femme enceinte sont paradoxalement fragmentaires. L'objectif de cette étude était de déterminer la séroprévalence de la toxoplasmose chez les femmes enceintes. Méthodes. Il s'agissait d'une étude transversale réalisée, à la maternité de l'Hôpital du District de Bossembelé, entre juin et septembre 2020. La population d'étude était constituée de femmes enceintes se présentant au laboratoire du District pour la sérologie toxoplasmique. Résultats. Au total, les données sérologiques de 50 femmes enceintes ont été analysées. L'âge moyen était de 25 ± 6 ans (extrème 16 et 40 ans). Les femmes au premier geste (n=20 soit 40 %) et les primipares (n= 30 soit 60 %) étaient prépondérantes. La sérologie était positive chez 15 patientes (30 %). Selon les caractéristiques sociodémographiques, la séroprévalence de la toxoplasmose était plus élevée chez les femmes de 20 à 35 ans (35,2 %), les femmes ayant été enceintes trois fois (88,8 %) et les femmes qui habitent le quartier Onoguia (66,66%). Les IgM étaient plus élevées chez les patients de la tranche d'âge de 20 à 35 ans (n=12), les femmes au 3e geste (n=8), les multipares (n=9) et chez celles habitant Bodoukpa (n=6). Les IgG étaient élevées chez les femmes enceintes de 20 à 35 ans (n=13), les femmes au 3e geste (n=7), les primipares (n=14) et celles habitant le quartier Bodoukpa (n=6). Parmi les patientes étudiées, 16 (32 %) étaient immunisées contre la toxoplasmose. Des 50 femmes, 4 avaient connu un avortement spontané durant les grossesses précédentes. Conclusion. Dans la présente étude, la séroprévalence de la toxoplasmose chez la femme enceinte est très fréquente. Une sensibilisation sur les risques de contamination, une surveillance sérologique systématique et des mesures d'hygiène devraient être proposées lors des consultations prénatales

Context and objective. Toxoplamosis is a ubiquitous anthropozoonosis that occupies a large place in human and veterinary medicine. The objective of the present study was to determine the seroprevalence of toxoplasmosis in pregnant women. Methods. This was a cross sectional study involving pregnant women presenting at the laboratory of the Bossembele District Hospital, Central African Republic between June and September 2020 for toxoplasmic serology. Results. A total of 50 pregnant women were examined. The age of patients varied from 16 to 40 years. The average age was 25 ± 6 years. Primigravida (n=20; or 40%) and primiparous women (n=30; or 60%) were more preponderant. Serology was positive in 15 patients (30 %). According to sociodemographic characteristics, the seroprevalence of toxoplasmosis was higher among women aged 20 to 35 (35.2 %), women who had been pregnant three times (88.8 %) and women who lived in the Onoguia neighborhood (66.6 %). IgM was higher in patients aged 20 to 35 years (n=12), in 3rd gravida women (n=8), in multiparous (n=9) and in those living in Bodoukpa (n=6). IgG was high in pregnant women aged 20 to 35 years (n=13), in 3rd gravida women (n=7), in primiparous women (n=14) and in those living in the Bodoukpa neighbourhood (n=6). Of the patients in the study, 16 turned out to be immune to toxoplasmosis. Among 50 women, 4 experienced spontaneous abortions during previous pregnancies. Conclusion. The seroprevalence of toxoplasmosis in the present study is common. Awareness on the risks of contamination, the systematic serological monitoring and the hygiene measures should be raised during antenatal consultations

Unexpected high frequency of unspecific reactivities by testing pre-epidemic blood specimens from Europe and Africa with SARS-CoV-2 IgG-IgM antibody rapid tests points to IgM as the Achilles heel.

We aimed to evaluate the rates of false-positive test results of three rapid diagnostic tests (RDTs) for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-specific immunoglobulin G (IgG) and IgM detection. Two serum panels from patients hospitalized in Paris, France, and from patients living in Bangui, Central African Republic, acquired before the 2019 COVID-19 outbreak, were tested by 3 CE IVD-labeled RDTs for SARS-CoV-2 serology (BIOSYNEX® COVID-19 BSS [IgG/IgM]; SIENNA™ COVID-19 IgG/IgM Rapid Test Cassette; NG-Test® IgG-IgM COVID-19). Detectable IgG or IgM reactivities could be observed in 31 (3.43%) of the 902 IgG and IgM bands of the 3 RDTs used with all pre-epidemic sera. The frequencies of IgG/IgM reactivities were similar for European (3.20%) and African (3.55%) sera. IgM reactivities were observed in 9 European and 14 African sera, while IgG reactivity was observed in only 1 African serum (15.1% vs. 0.66%). The test NG-Test® IgG-IgM COVID-19 showed the highest rates of IgG or IgM reactivities (6.12% [18/294]), while the test BIOSYNEX® COVID-19 BSS (IgG/IgM) showed the lowest rate (1.36% [4/294]). Some combinations of 2 RDTs in series allowed decreasing significantly the risk of false-positive test results. Our observations point to the risk of false-positive reactivities when using currently available RDT for SARS-CoV-2 serological screening, especially for the IgM band, even if the test is CE IVD-labeled and approved by national health authorities, and provide the rational basis for confirmatory testing by another RDT in case of positive initial screening.

Escalating and sustained immunovirological dissociation among antiretroviral drug-experienced perinatally human immunodeficiency virus-1-infected children and adolescents living in the Central African Republic: A STROBE-compliant study.

Sub-Saharan Africa has the vast majority (∼90%) of new pediatric acquired immunodeficiency syndrome cases worldwide. Biologically monitoring HIV-infected pediatric populations remains challenging. The differential interest of human immunodeficiency virus (HIV)-1 RNA loads and CD4 T-cell counts is debated for the treatment of pediatric acquired immunodeficiency syndrome patients.Long-term antiretroviral treatment (ART) outcomes regarding immunological and virological surrogate markers were longitudinally evaluated between 2009 and 2014 (over 57 months) in 245 perinatally HIV-1-infected children and adolescents born from HIV-infected mothers, treated at inclusion for at least 6 months by the World Health Organization-recommended ART in Bangui, Central African Republic.Patients were monitored over time biologically for CD4 T-cell counts, HIV-1 RNA loads, and drug resistance mutation genotyping.Children lost to follow-up totaled 6%. Four categories of immunovirological responses to ART were observed. At baseline, therapeutic success with sustained immunological and virological responses was observed in 80 (32.6%) children; immunological and virologic nonresponses occurred in 32 (13.0%) children; finally, the majority (133; 54.2%) of the remaining children showed discordant immunovirological responses. Among them, 33 (13.4%) children showed rapid virological responses to ART with an undetectable viral load, whereas immunological responses remained absent after 6 months of treatment and increased progressively over time in most of the cases, suggesting slow immunorestoration. Notably, nearly half of the children (40.8% at baseline and 48.2% at follow-up) harbored discordant immunovirological responses with a paradoxically high CD4 T-cell count and HIV-1 RNA load, which are always associated with high levels of drug resistance mutations. The latter category showed a significant increase over time, with a growth rate of 1.23% per year of follow-up.Our STROBE-compliant study demonstrates the high heterogeneity of biological responses under ART in children with frequent passage from 1 category to another over time. Close biological evaluation with access to routine plasma HIV-1 RNA load monitoring is crucial for adapting the complex outcomes of ART in HIV-infected children born from infected mothers.

Purifying Selection in Human Immunodeficiency Virus-1 pol Gene in Perinatally Human Immunodeficiency Virus-1-Infected Children Harboring Discordant Immunological Response and Virological Nonresponse to Long-Term Antiretroviral Therapy.

BACKGROUND: Biological monitoring of antiretroviral treatment (ART) in human immunodeficiency virus (HIV)-infected pediatric population remains challenging. The aim of the present study was to assess the long-term HIV-1 genetic diversity in pol gene in HIV-1-infected children in virological failure under antiretroviral regimen adapted according to the successive World Health Organization (WHO) guidelines for resource-constrained settings. METHODS: HIV-1 diversity in pol gene was assessed in HIV-1-infected children and adolescents born from HIV-infected mothers (median age at follow-up: 13.8 years) in virological failure (VF+) despite long-term regimen recommended by the WHO. The numbers of nonsynonymous substitutions per potential nonsynonymous site (dN) and of synonymous substitutions at potential synonymous sites (dS) in HIV-1 pol gene and the dN/dS ratios were used to estimate the selective pressure on circulating HIV-1. RESULTS: The immunological responses to ART basically corresponded to: 1) Full therapeutic failure with immunological (I-) and virological nonresponses in one-quarter (24.6%) of study children ((I-, VF+) subgroup); 2) Discordant immunovirological responses with paradoxical high CD4 T cell counts (I+) and high HIV-1 RNA load in the remaining cohort patients (75.4%) ((I+, VF+) subgroup). The mean dS was 1.8-fold higher in (I+, VF+) than (I-, VF+) subgroup (25.9 ± 18.4 vs. 14.3 ± 10.8). In the (I+, VF+) subgroup, the mean dS was 1.6-fold higher than the mean dN. Finally, the mean dN/dS ratio was 2.1-fold lower in (I+, VF+) than (I-, VF+) subgroup (0.6 ± 0.3 vs. 1.3 ± 0.7), indicating purifying selection in the immunovirological discordant (I+, VF+) subgroup and positive selection in the immunovirological failure (I-, VF+) subgroup. CONCLUSIONS: Children and adolescents in immunovirological therapeutic failure harbor positive selection of HIV-1 strains favoring diversifying in pol-encoded amino acids. In contrast, children with persistent discordant immunovirological responses show accumulation of mutations and purifying selection in pol gene sequences, indicating limited genetic evolution and likely suggesting genetic adaptation of viruses to host functional constraints.

Infrequent detection of human papillomavirus infection in head and neck cancers in the Central African Republic: a retrospective study.

We carried out a retrospective study on the prevalence of HPV and genotype distribution by nested PCR and nucleotide sequencing analysis, in formalin-fixed, paraffin-embedded biopsies of 135 head and neck cancers (HNC) and 29 cervical cancers received between 2009 and 2017 for diagnosis at the Laboratoire National de Biologie Clinique et de Santé Publique of Bangui, the capital city of the Central African Republic. One oropharyngeal squamous cell carcinoma sample was positive for HPV type 16. The overall HPV prevalence in HNC biopsies was 0.74% (95% CI: 0.0-2.2). Among the 29 cervical cancer samples, 19 (65.5%; 95% CI: 48.2-82.8) were positive for HPV. These results indicate that HNC are infrequently associated with HPV infection in the Central African Republic.

High predictive efficacy of integrase strand transfer inhibitors in perinatally HIV-1-infected African children in therapeutic failure of first- and second-line antiretroviral drug regimens recommended by the WHO.

OBJECTIVES: The predictive efficacy of integrase (IN) strand transfer inhibitors (INSTIs) was investigated in HIV-infected children born to HIV-infected mothers in Africa. METHODS: Plasma was collected at the Complexe Pédiatrique of Bangui, Central African Republic, from INSTI-naive children (n = 8) and adolescents (n = 10) in virological failure (viral load >1000 copies/mL) after 5 years of first- and/or second-line combination ART (cART). IN, reverse transcriptase (RT) and protease (P) genes were genotyped and drug resistance mutations (DRMs) to INSTIs, NRTIs, NNRTIs and PIs were interpreted using the Stanford algorithm. RESULTS: Successful IN, RT and P genotypes were obtained for 18, 13 and 15 children (median age 11 years, range 5-18; 8 were female), respectively. Two (2/18; 11.1%) viruses from children treated with a first-line regimen had INSTI DRMs at codon 138 (E138K and E138T), which is known to harbour major resistance mutations, and also had the accessory mutations L74I, G140K, G140R and G163R. The majority (16/18; 88.9%) of HIV-1 IN sequences demonstrated full susceptibility to all major INSTIs with a high frequency of natural polymorphic mutations. Most (12/15; 80%) genotyped viruses harboured at least one major DRM conferring resistance to at least one of the WHO-recommended antiretroviral drugs (NNRTIs, NRTIs and PIs) prescribed in first- and second-line regimens. CONCLUSIONS: INSTIs could be proposed in first-line regimens in the majority of African children or adolescents and may constitute relevant therapeutic alternatives as second- and third-line cART regimens in HIV-infected children and adolescents living in sub-Saharan Africa.

Usefulness of simultaneous screening for HIV-specific and HCV-specific antibodies and HBsAg by a capillary-based multiplex rapid diagnostic test to strengthen linkage-to-care in sub-Saharan patients attending sexually transmitted infection clinic.

Adult outpatients attending the main sexually transmitted infection clinic of Bangui, Central African Republic, were prospectively subjected to a multiplex rapid diagnostic test for human immunodeficiency virus (HIV), hepatitis B virus (HBV), and hepatitis C virus (HCV). In group I (n = 208) of patients already followed for HIV, 6 (2.9%) were unexpectedly negative, thus corresponding to false positive for HIV by the national HIV algorithm; hepatitis B surface antigen and HCV positivities were high (18.7% and 4.3%, respectively). In group II (n = 71) of patients with unknown HIV status, at least 1 chronic viral disease was diagnosed in 26 (36.6%) patients, including 5 (7.1%) HIV, 17 (23.9%) HBV, and 3 (4.2%) HCV infections.

Unusual and unique distribution of anal high-risk human papillomavirus (HR-HPV) among men who have sex with men living in the Central African Republic.

BACKGROUND: High-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. METHODS: MSM 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. RESULTS: 42 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). CONCLUSION: MSM 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.

Task-shifting of CD4 T cell count monitoring by the touchscreen-based Muse™ Auto CD4/CD4% single-platform system for CD4 T cell numeration: Implication for decentralization in resource-constrained settings.

BACKGROUND: The accuracy of CD4 T cell monitoring by the recently developed flow cytometry-based CD4 T cell counting Muse™ Auto CD4/CD4% Assay analyzer (EMD Millipore Corporation, Merck Life Sciences, KGaA, Darmstadt, Germany) was evaluated in trained lay providers against laboratory technicians. METHODS: After 2 days of training on the Muse™ Auto CD4/CD4% analyzer, EDTA-blood samples from 6 HIV-positive and 4 HIV-negative individuals were used for CD4 T cell counting in triplicate in parallel by 12 trained lay providers as compared to 10 lab technicians. RESULTS: Mean number of CD4 T cells in absolute number was 829 ±â€¯380 cells/µl by lay providers and 794 ±â€¯409 cells/µl by technicians (P > 0.05); and in percentage 36.2 ±â€¯14.8%CD4 by lay providers and 36.1 ±â€¯15.0%CD4 by laboratory technician (P > 0.05). The unweighted linear regression and Passing-Bablok regression analyses on CD4 T cell results expressed in absolute count revealed moderate correlation between CD4 T cell counts obtained by lay providers and lab technicians. The mean absolute bias measured by Bland-Altman analysis between CD4 T cell/µl obtained by lay providers and lab technicians was -3.41 cells/µl. Intra-assay coefficient of variance (CV) of Muse™ Auto CD4/CD4% in absolute number was 10.1% by lay providers and 8.5% by lab technicians (P > 0.05), and in percentage 5.5% by lay providers and 4.4% by lab technicians (P > 0.05). The inter-assay CV of Muse™ Auto CD4/CD4% in absolute number was 13.4% by lay providers and 10.3% by lab technicians (P > 0.05), and in percentage 7.8% by lay providers and 6.9% by lab technicians (P > 0.05). CONCLUSIONS: The study demonstrates the feasibility of CD4 T cell counting using the alternative flow cytometer Muse™ Auto CD4/CD4% analyzer by trained lay providers and therefore the practical possibility of decentralization CD4 T cell counting to health community centers.

High levels of virological failure with major genotypic resistance mutations in HIV-1-infected children after 5 years of care according to WHO-recommended 1st-line and 2nd-line antiretroviral regimens in the Central African Republic: A cross-sectional study.

A large cohort of 220 HIV-1-infected children (median [range] age: 12 [4-17] years) was cared and followed up in the Central African Republic, including 198 in 1st-line and 22 in 2nd-line antiretroviral regimens. Patients were monitored clinically and biologically for HIV-1 RNA load and drug resistance mutations (DRMs) genotyping. A total of 87 (40%) study children were virological responders and 133 (60%) nonresponders. In children with detectable viral load, the majority (129; 97%) represented a virological failure. In children receiving 1st-line regimens in virological failure for whom genotypic resistance test was available, 45% displayed viruses harboring at least 1 DRM to NNRTI or NRTI, and 26% showed at least 1 major DRM to NNRTI or NRTI; more than half of children in 1st-line regimens were resistant to 1st-generation NNRTI and 24% of the children in 1st-line regimens had a major DRMs to PI. Virological failure and selection of DRMs were both associated with poor adherence. These observations demonstrate high rate of virological failure after 3 to 5 years of 1st-line or 2nd-line antiretroviral treatment, which is generally associated with DRMs and therapeutic failure. Overall, more than half (55%) of children receiving 1st-line antiretroviral treatment for a median of 3.4 years showed virological failure and antiretroviral-resistance and thus eligible to 2nd-line treatment. Furthermore, two-third (64%) of children under 2nd-line therapy were eligible to 3rd-line regimen. Taken together, these observations point the necessity to monitor antiretroviral-treated children by plasma HIV-1 RNA load to diagnose as early as possible the therapeutic failure and operate switch to a new therapeutic line.

Performance evaluation of the touchscreen-based Muse™ Auto CD4/CD4% single-platform system for CD4 T cell numeration in absolute number and in percentage using blood samples from children and adult patients living in the Central African Republic.

BACKGROUND: The new microcapillary and fluorescence-based EC IVD-qualified Muse™ Auto CD4/CD4% single-platform assay (EMD Millipore Corporation, Merck Life Sciences, KGaA, Darmstadt, Germany) for CD4 T cell numeration in absolute number and in percentage was evaluated using Central African patients' samples compared against the reference EC IVD-qualified BD FACSCount (Becton-Dickinson, USA) flow cytometer. METHODS: EDTA-blood samples from 124 adults, 10 adolescents, 13 children and 3 infants were tested in parallel at 2 reference laboratories in Bangui. RESULTS: The Muse™ technique was highly reproducible, with low intra- and inter-run variabilities less than 15%. CD4 T cell counts of Muse™ and BD FACSCount in absolute number and percentage were highly correlated (r2 = 0.99 and 0.98, respectively). The mean absolute bias between Muse™ and BD FACSCount cells in absolute number and percentage were -5.91 cells/µl (95% CI -20.90 to 9.08) with limits of agreement from -77.50 to 202.40 cells/µl, and +1.69 %CD4 (95% CI ±1.29 to +2.09), respectively. The percentages of outliers outside the limits of agreement were nearly similar in absolute number (8%) and percentage (10%). CD4 T cell counting by Muse™ allowed identifying the majority of individuals with CD4 T cell <200, <350 or <750 cells/µl corresponding to the relevant thresholds of therapeutic care, with sensitivities of 95.5-100% and specificities of 83.9-100%. CONCLUSIONS: The Muse™ Auto CD4/CD4% Assay analyzer is a reliable alternative flow cytometer for CD4 T lymphocyte enumeration to be used in routine immunological monitoring according to World Health Organization recommendations in HIV-infected adults as well as children living in resource-constrained settings.

Analytical Performances of Human Immunodeficiency Virus Type 1 RNA-Based Amplix® Real-Time PCR Platform for HIV-1 RNA Quantification.

Objectives. We evaluated the performances of Amplix real-time PCR platform developed by Biosynex (Strasbourg, France), combining automated station extraction (Amplix station 16 Dx) and real-time PCR (Amplix NG), for quantifying plasma HIV-1 RNA by lyophilized HIV-1 RNA-based Amplix reagents targeting gag and LTR, using samples from HIV-1-infected adults from Central African Republic. Results. Amplix real-time PCR assay showed low limit of detection (28 copies/mL), across wide dynamic range (1.4-10 log copies/mL), 100% sensitivity and 99% specificity, high reproducibility, and accuracy with mean bias < 5%. The assay showed excellent correlations and concordance of 95.3% with the reference HIV-1 RNA load assay (Roche), with mean absolute bias of +0.097 log copies/mL by Bland-Altman analysis. The assay was able to detect and quantify the most prevalent HIV-1 subtype strains and the majority of non-B subtypes, CRFs of HIV-1 group M, and HIV-1 groups N and O circulating in Central Africa. The Amplix assay showed 100% sensitivity and 99.6% specificity to diagnose virological failure in clinical samples from antiretroviral drug-experienced patients. Conclusions. The HIV-1 RNA-based Amplix real-time PCR platform constitutes sensitive and reliable system for clinical monitoring of HIV-1 RNA load in HIV-1-infected children and adults, particularly adapted to intermediate laboratory facilities in sub-Saharan Africa.

Antimicrobial Resistance of Enteric Salmonella in Bangui, Central African Republic.

Introduction. The number of Salmonella isolated from clinical samples that are resistant to multiple antibiotics has increased worldwide. The aim of this study was to determine the prevalence of resistant Salmonella enterica isolated in Bangui. Methods. All enteric Salmonella strains isolated from patients in 2008 were identified and serotyped, and the phenotypes of resistance were determined by using the disk diffusion method. Nine resistance-associated genes, bla TEM , bla OXA , bla SHV , tetA, aadA1, catA1, dhfrA1, sul I, and sul II, were sought by genic amplification in seven S.e. Typhimurium strains. Results. The 94 strains isolated consisted of 47 S.e. Typhimurium (50%), 21 S.e. Stanleyville (22%), 18 S.e. Enteritidis (19%), 4 S.e. Dublin (4%), 4 S.e. Hadar (4%), and 1 S.e. Papuana (1%). Twenty-five (28%) were multiresistant, including 20 of the Typhimurium serovar (80%). Two main phenotypes of resistance were found: four antibiotics (56%) and to five antibiotics (40%). One S.e. Typhimurium isolate produced an extended-spectrum ß-lactamase (ESBL). Only seven strains of S.e. Typhimurium could be amplified genically. Only phenotypic resistance to tetracycline and aminosides was found. Conclusion. S. Typhimurium is the predominant serovar of enteric S. enterica and is the most widely resistant. The search for resistance genes showed heterogeneity of the circulating strains.