Evaluation of the Determine™ fourth generation HIV rapid assay

Assays that detect p24 antigen reduce the diagnostic window period of HIV testing. Most point-of-care HIV assays have poor sensitivity to diagnose acute HIV infection as they only detect antibodies against HIV-1 and HIV-2 (HIV-1/2). This was a cross-sectional laboratory-based study that evaluated the performance of the DetermineTM HIV-1/2 Ag/Ab Combo fourth generation rapid strip – currently the only rapid assay that detects both HIV-1/2 antibodies and p24 antigen. A total of 79 serum specimens (29 positive for HIV antibodies only, 14 positive for HIV antibodies and p24 antigen, 20 HIV-negative, and 16 positive for p24 antigen only) were used for the evaluation. Results were compared with those from validated fourth generation HIV ELISAs. The DetermineTM Combo rapid strips had a sensitivity of 90.7% and a specificity of 100% for the detection of HIV-1/2 antibodies. Its sensitivity for the detection of p24 antigen was only 10% (3 out of 30 p24 antigen positive specimens). This implies that most acute HIV infections will be missed with this assay. The need for a point-of-care assay which can detect acute HIV infection reliably still remains, particularly for use in a high prevalence setting such as South Africa.The authors would like to thank the South African Regional Headquarters of Alere (Waltham, USA) for providing DetermineTM HIV-1/2 Ag/Ab Combo kits for validation purposes, as well as funding provided for the purchase of the seroconversion panel from the South African National Blood Service.http:// www.elsevier.com/locate/jviromethb2013ay201

HIV-1/2 differentiation in a South African public laboratory

BACKGROUND : The human immunodeficiency virus type-2 (HIV-2) prevalence in South Africa (SA) is unknown, however, sporadic cases have been reported. Human immunodeficiency virus -1 and 2 differentiation is not part of most South African public laboratories’ testing algorithm. Human immunodeficiency virus -2 diagnosis using serology assays may be complicated by HIV-1 and HIV-2 antibody cross-reactivity. OBJECTIVES : To determine the proportion of HIV-2 infections in specimens that tested HIV-1/2 positive at a public laboratory in Tshwane. METHOD: A total of 480 specimens that were previously tested with fourth generation ELISA platforms (Modular E170 [Roche, Switzerland] and Architect i2000 [Abbott, Germany]) were randomly selected. Human immunodeficiency virus -1 and 2 antibody differentiation testing was carried out using the Multispot HIV-1/2 rapid assay (Bio-Rad Laboratories, USA). An inhouse nested HIV-2 PCR assay targeting the 5'-long terminal repeats (5'-LTR) region was evaluated and used as a confirmatory test. RESULT: The study tested 480 HIV-1/2 seropositive patients and their mean age was 36.7 years (range 3–82 years). Of the 480 patients, 292 (60.8%) were female, 182 (37.9%) were male and 6 (1.3%) were not specified. Human immunodeficiency virus differentiation results were as follows: 466 (97.1%) were positive for only HIV-1 antibodies, 11 (2.3%) [95%CI: (0.98%; 3.74%)] were positive for both HIV-1 and HIV-2 antibodies, 3 (0.6%) were negative for both antibodies and none were positive for only HIV-2 antibodies. Of the 11 specimens with both HIV-1 and HIV-2 antibodies, seven had sufficient volume for confirmatory testing and were all negative on the in-house HIV-2 PCR assay. CONCLUSION: The multispot HIV-1/2 rapid assay demonstrated cross-reactivity between HIV-1 and HIV-2 antibodies. Human immunodeficiency virus -2 infections were not detected.NHLS Research Trust; Discovery Foundation and University of Pretoria Research grant.http://www.sajhivmed.org.za/index.php/hivmeddm2022Medical Virolog

Antenatal screening for hepatitis B virus in HIV-infected and uninfected pregnant women in the Tshwane district of South Africa

BACKGROUND. Despite enormous strides in preventing hepatitis B virus (HBV) infection, perinatal transmission still contributes significantly to HBV epidemiology worldwide; this could account for approximately 50% of chronically infected individuals. OBJECTIVE. To assess the need for HBV screening in antenatal clinics in the HIV/AIDS era. METHODS. This was a retrospective study conducted at the antenatal clinic of 1 Military Hospital, Tshwane, South Africa. Laboratory data for HBV, HIV and CD4 count were obtained and analysed for the period January 2008 - December 2013. RESULTS. A total of 2 513 patients’ results were retrieved and 2 368 patients were enrolled as both their HBV and HIV serology results were available. The mean age of participants was 29 years (range 14 - 46). HIV prevalence in this study was 20.5% (95% confidence interval (CI) 0.189 - 0.222). The median CD4 count in HIV-infected patients was 522 cells/μL (interquartile range 370 - 711). There was an overall HBV prevalence of 0.8% (95% CI 0.005 - 0.011). The hepatitis B surface antigen (HBsAg) prevalence was significantly higher (2.1%) among HIV co-infected compared with HIV-uninfected patients (0.4%) (p=0.0001). Hepatitis e antigen (HBeAg) positivity was 30% in the HIV co-infected compared with 37.6% in the HIV-uninfected individuals (p=0.7400). CONCLUSION. This study showed a significantly higher HBV prevalence in HIV-infected compared with HIV-uninfected patients. The comparable HBeAg prevalence between the two groups indicates that both were at an increased risk of vertical transmission, therefore demonstrating a need for antenatal screening for HBV. Since antenatal screening is often not affordable in low-income countries, administration of HBV vaccine at birth is needed for prevention of vertical transmission.http://www.samj.org.zaam2016Medical VirologyObstetrics and Gynaecolog

Variability of the preC/C region of hepatitis B virus genotype A from a South African cohort predominantly infected with HIV

ABSTRACT Hepatitis B virus (HBV) is a serious global health problem, and HBV genotype is an important determinant of disease progression and treatment outcome. The aim of this study was to assess variations of the precore/core (preC/C) region in HBV genotype A. Sequencing of the preC/C and surface (S) genes of HBV was performed on plasma samples from 20 HBV/HIV co-infected and 5 HBV mono-infected individuals. All preC/C study sequences clustered with subgenotype A1, except for 2 which clustered with subgenotype D4 reference strains. The nucleotide and amino acid variability was far higher in the preC/C region than in the S region. Mutations associated with reduction or failure of HBV e-antigen (HBeAg) production were observed, with a preC start codon mutation being common (24%). Other mutations (e.g. P5H/L and I97L) associated with severe liver disease were also noticed, some of which were located in the major histocompatibility restricted sites. PreC/C intergenotype nucleotide divergence was >7%, while subgenotypes differed by 2.5 -7%. Several study sequences were highly divergent from other African subgenotype A1 strains. This study showed that HBeAg-negative chronic hepatitis B is underestimated in subgenotype A1, and also highlighted the variant South African A1 strains. The major advantage of preC/C sequencing is that it informs patient management as HBeAgnegative chronic hepatitis B responds poorly to conventional interferon-α therapy, and some guidelines treat HBeAg-negative chronic hepatitis B differently from HBeAg-positive chronic hepatitis B. These data suggest that subgenotype A1 may be more involved in severe HBVrelated diseases

Pre-treatment minority HIV-1 drug resistance mutations and long term virological outcomes : is prediction possible?

BACKGROUND : Although the use of highly active antiretroviral therapy in HIV positive individuals has proved to be effective in suppressing the virus to below detection limits of commonly used assays, virological failure associated with drug resistance is still a major challenge in some settings. The prevalence and effect of pre-treatment resistance associated variants on virological outcomes may also be underestimated because of reliance on conventional population sequencing data which excludes minority species. We investigated long term virological outcomes and the prevalence and pattern of pre-treatment minority drug resistance mutations in individuals initiating HAART at a local HIV clinic. METHODS : Patient’s records of viral load results and CD4 cell counts from routine treatment monitoring were used and additional pre-treatment blood samples for Sanger sequencing were obtained. A selection of pretreatment samples from individuals who experienced virological failure were evaluated for minority resistance associated mutations to 1 % prevalence and compared to individuals who achieved viral suppression. RESULTS : At least one viral load result after 6 months or more of treatment was available for 65 out of 78 individuals followed for up to 33 months. Twenty (30.8 %) of the 65 individuals had detectable viremia and eight (12.3 %) of them had virological failure (viral load > 1000 RNA copies/ml) after at least 6 months of HAART. Viral suppression, achieved by month 8 to month 13, was followed by low level viremia in 10.8 % of patients and virological failure in one patient after month 20. There was potentially reduced activity to Emtricitabine or Tenofovir in three out of the eight cases in which minority drug resistance associated variants were investigated but detectable viremia occurred in one of these cases while the activity of Efavirenz was generally reduced in all the eight cases. CONCLUSIONS : Early viral suppression was followed by low level viremia for some patients which may be an indication of failure to sustain viral suppression over time. The low level viremia may also be representing early stages of resistance development. The mutation patterns detected in the minority variants showed potential reduced drug sensitivity which highlights their potential to dominate after treatment initiation. TRIAL REGISTRATION : Not applicable.Additional file 1: Figure S1. Deep sequencing coverage. C – E shows sequencing coverage for samples with virologicalfailure (L031, L054 and L064 respectively), F shows coverage for a sample with detectable viremia (L009)and G and H show coverage for virally suppressed samples (L074 and L075 respectively). Mutations wereexcluded from analysis for any of the following: noisy mutations filtering, coverage filtering, forward/ reverse unbalanced frequency and forward/reverse unbalanced coverage.This work is based on the research supported by grants awards from South African Research Chairs Initiative of the Department of Science and Technology and National Research Foundation of South Africa (C.T. Tiemessen), the HIV Research Trust (M.L. Mzingwane), the National Health Laboratory Service Research Trust and the Poliomyelitis Research Foundation.http://www.virologyj.comam2016Medical Virolog

Hand-foot-and-mouth disease caused by coxsackievirus A6 in a patient infected with HIV

Hand, foot and mouth disease (HFMD) is common in children ≤ 5 years of age, and is mainly caused by enterovirus 71 and coxsackievirus A16 (CVA6). A 12-year-old boy on treatment for human immunodeficiency virus (HIV) presented to an HIV clinic with fever and a rash on the palms and soles. The syphilis test were negative. Enterovirus was identified from a stool sample by PCR and characterised as as coxsackievirus A6(CVA6). The patient completely recovered a week later. CVA6 has recently been associated with HFMD. This case highlights the significance of the laboratory confirmation of suspected HFMD cases aand phylogenetic analysis of the identified virus.http://www.sajei.co.za/index.php/SAJEIam201

Evaluation of COVID-19 impact on early infant diagnosis of human immune defiency virus

https://drive.google.com/file/d/15cjp0gBKQJFSMljIdOgqn5OLObKwnK_v/view?usp=sharinghttps://drive.google.com/drive/folders/1X8gKZuWAt18-Ob8ozv54RgVpSuCOLWa7?usp=sharinghttps://drive.google.com/drive/folders/1ajSLECaa0MzNFZQdbFa5YL915LYuzonH?usp=sharin

Variability of the preC/C region of hepatitis B virus genotype A from a South African cohort predominantly infected with HIV

Hepatitis B virus (HBV) is a serious global health problem, and HBV genotype is an important determinant of disease progression and treatment outcome. The aim of this study was to assess variations of the precore/core (preC/C) region in HBV genotype A. Sequencing of the preC/C and surface (S) genes of HBV was performed on plasma samples from 20 HBV/HIV co-infected and 5 HBV mono-infected individuals. All preC/C study sequences clustered with subgenotype A1, except for two which clustered with subgenotype D4 reference strains. The nucleotide and amino acid variability was far higher in the preC/C region than in the S region. Mutations associated with reduction or failure of HBV e-antigen (HBeAg) production were observed, with a preC start codon mutation being common (24%). Other mutations (e.g., P5H/L and I97L) associated with severe liver disease were also noticed, some of which were located in the major histocompatibility restricted sites. PreC/C intergenotype nucleotide divergence was >7%, while subgenotypes differed by 2.5–7%. Several study sequences were highly divergent from other African subgenotype A1 strains. This study showed that HBeAg-negative chronic hepatitis B is underestimated in subgenotype A1, and also highlighted the variant South African A1 strains. The major advantage of preC/C sequencing is that it informs patient management as HBeAg-negative chronic hepatitis B responds poorly to conventional interferon-α therapy, and some guidelines treat HBeAg-negative chronic hepatitis B differently from HBeAg-positive chronic hepatitis B. These data suggest that subgenotype A1 may be more involved in severe HBV-related diseaseshttp://onlinelibrary.wiley.com/journal/10.1002/(ISSN)1096-9071/hb201

HBV/HIV co-infection: The dynamics of HBV in South African patients with AIDS

OBJECTIVE: As sub-Saharan Africa is highly endemic for hepatitis B virus (HBV) and human immunodeficiency virus (HIV) infections, and their co-infection requires special management, we aimed to assess the serological and molecular characteristics of HBV in patients with AIDS. DESIGN: This was a cross-sectional, case control study, which enrolled 200 patients with AIDS and 200 HIV-negative controls. HBV serology was done in all participants and HCV serology in participants with a hepatitis B core antibody (anti-HBc) only serological pattern. Nested HBV polymerase chain reaction (PCR) and HBV viral load assays were used for HBV molecular detection. RESULTS: Hepatitis B surface antigen (HBsAg) prevalence was 3-fold higher while the 'anti-HBc only' pattern was 6-fold higher in the AIDS group compared with the controls. Mean HBV viral load was significantly higher in HBsAg-positive patients with CD4+ cell counts <100 cells/μl than in patients with CD4+ cell counts of 100-200 cells/μl (p=0.019). There were markedly reduced hepatitis B surface antibody (anti-HBs) titres in the AIDS group compared with the controls (p=0.002). A significant proportion of AIDS patients with an 'anti-HBc only' pattern had CD4+ cell counts <100 cells/μl (p=0.004). Occult HBV prevalence was 3.5% in the AIDS group compared with 1% in the controls (p=0.092). When occult HBV infection was taken into consideration, the overall HBV prevalence became 10% in the AIDS group and 3% in the control group. CONCLUSION: We showed an increased HBV prevalence in patients with AIDS and identified a CD4+ cell count <100 cells/μl as a major risk factor for the 'anti-HBc only' pattern and increased HBV replication. These data have significant public health implications for HBV in developing countries, especially in areas where antiretroviral (ARV) guidelines do not cater for HBV / HIV co-infection.The National Health Laboratory Service (NHLS) and Poliomyelitis Research Foundation (PRF).http://www.samj.org.z

Low seroprevalence of hepatitis E virus in pregnant women in an urban area near Pretoria, South Africa

OBJECTIVES : Hepatitis E virus (HEV) infection is a globally neglected health problem with a high burden in resource-poor communities. Pregnant women are at increased risk of complications. This pilot study sought to assess the seroprevalence of HEV infection in pregnant women at Dr George Mukhari Academic Hospital, South Africa. METHODS : Stored serum samples from 384 HIV-uninfected pregnant women attending the antenatal clinic were initially screened for HEV total antibody. Positive samples were further evaluated for the presence of IgG and IgM antibody isotypes, using commercial ELISA assays. HEV RNA was assessed in antibody-positive samples utilizing qRT-PCR assay. RESULTS : The sample consisted of women with a median age of 31 years (interquartile range: 28–35 years). Total HEV antibody was detected in 12/384 (3.13%, 95% CI: 1.80–5.38) of these pregnant women. All 12 samples were IgG HEV antibody positive, but none tested positive for IgM antibody or for HEV RNA, demonstrating a lack of current or recent exposure. CONCLUSIONS : Our study revealed a low seroprevalence of HEV among pregnant women from an urban area north of Pretoria. This observation warrants further attention to the circulation of HEV in this population, and a greater understanding of the epidemiology of the infection in South Africa.The South African Ministry of Higher Education, Science and Innovation, and the National Research Foundation.http://www.elsevier.com/locate/ijregihj2023Medical Virolog