Justen Manasa
University of Zimbabwe, Medical Microbiology, Faculty Member
- HIV/AIDS, Molecular Genetics, Phylogenetics, Infectious Diseases, Virology, Human Genetics, and 21 moreMolecular virology, Sequence Analysis, HIV medicine, Hepatitis C, Virus-Host Interactions, RNA viruses, Molecular hematology, Meta-analysis and systematic review (Health Sciences), DNA viruses, Retroviruses, Proteome-informatics, HTLV (Retroviruses), Biotechnology, Bioinformatics, Molecular Biology, Genetics, Immunology, Open Source Software, Genome Wide Association Studies (GWAS), HIV drug resistance, and Molecular Epidemiologyedit
- I am a molecular virologist with interest in viral evolution and drug resistance. The focus of my research is currently HIV-1 Subtype C drug resistance in Southern Africa.edit
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Introduction: pre-treatment drug resistance (PDR) can compromise the 3rd 95-95-95 global target for viral load suppression. The high complexity and cost of genotyping assays limits routine testing in many resource limited settings (RLS).... more
Introduction: pre-treatment drug resistance (PDR) can compromise the 3rd 95-95-95 global target for viral load suppression. The high complexity and cost of genotyping assays limits routine testing in many resource limited settings (RLS). We assessed the performance of a rapid HIV-1 drug resistance assay, the Pan Degenerate Amplification and Adaptation (PANDAA) assay when screening for significant HIV-1 drug resistance mutations (DRMs) such as K65R, K103NS, M184VI, Y181C and G190A. Methods: we used previously generated amplicons from a cross-sectional study conducted between October 2018 and February 2020 of HIV-1 infected antiretroviral therapy (ART)-naïve or those reinitiating 1st line ART (18 years or older). The performance of the PANDAA assay in screening K65R, K103NS, M184VI, Y181C, and G190A mutations compared to the reference assay, Sanger sequencing was evaluated by Cohen´s kappa coefficient on Stata version 14 (StataCorp LP, College Station, TX, USA). Results: one hundred and twenty samples previously characterized by Sanger sequencing were assessed using PANDAA. PDR was found in 14% (17/120). PDR to non-nucleoside reverse transcriptase inhibitors (NNRTIs) was higher at 13% (16/120) than PDR to nucleotide reverse transcriptase inhibitors (NRTIs), 3% (3/120). The PANDAA assay showed a strong agreement with the reference assay, i.e. Sanger sequencing for all five target DRMs (kappa (95%CI); 0.93 (0.78-0.98)) and NNRTI DRMs (kappa (95%CI); 0.93 (0.77-0.980), and a perfect agreement for NRTI DRMs (kappa (95%CI); 1.00 (0.54-1.00)). Conclusion: the PANDAA assay is a simple and rapid method to identify significant HIV DRMs in plasma samples as an alternative to Sanger sequencing in many RLS.
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Objectives:The integrase strand inhibitor dolutegravir (DTG) combined with tenofovir and lamivudine (TLD) is a single tablet regimen recommended for 1st, 2nd and 3rd line public health ART. We determined drug resistance mutations (DRMs)... more
Objectives:The integrase strand inhibitor dolutegravir (DTG) combined with tenofovir and lamivudine (TLD) is a single tablet regimen recommended for 1st, 2nd and 3rd line public health ART. We determined drug resistance mutations (DRMs) and evaluated the predictive efficacy of a TLD containing regimen for viremic adolescents and young adults in Harare, Zimbabwe.Methods:We sequenced plasma viral RNA from HIV-1 infected adolescents and young adults on 1st and 2nd line ART with confirmed virologic failure (VL>1000 copies/ml) and calculated genotypic susceptibility scores (tGSS) to current 2nd, 3rd line and DTG regimens.Results:160 participants were genotyped; 112 (70%) on 1st line and 48 (30%) on 2nd line, median (IQR) age 18 (15-19) and duration of ART(IQR) was 6 (4-8) years. Major DRMs were present in 94% and 67% of 1st and 2nd line failures respectively (p<0.001). Dual class resistance to NRTIs and NNRTIs was detected in 96 (60%) of 1st line failures; PI DRMs were detected in a minority (10%) of 2nd line failures. A total genotypic susceptibility score (tGSS) ≤2 may risk PI or DTG monotherapy in 11% and 42% of 1st line failures switching to 2nd line PIs and TLD respectively.Conclusion:Among adolescents and young adults, current PI based 2nd line therapies are poorly tolerated, more expensive and adherence is poor. In 1st line failure, implementation of TLD for many adolescents and young adults on long-term ART may require additional active drug(s). Drug resistance surveillance and susceptibility scores may inform strategies for the implementation of TLD.
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HIV drug resistance (HIVDR) is a barrier to sustained virologic suppression in low- and middle-income countries (LMICs). Point mutation assays targeting priority drug resistance mutations (DRMs) are being evaluated to improve access to... more
HIV drug resistance (HIVDR) is a barrier to sustained virologic suppression in low- and middle-income countries (LMICs). Point mutation assays targeting priority drug resistance mutations (DRMs) are being evaluated to improve access to HIVDR testing. In a cross-sectional study (June 2018 to September 2019), we evaluated the diagnostic accuracy of a simple and rapid HIVDR assay (the pan-degenerate amplification and adaptation [PANDAA] assay targeting the mutations K65R, K103NS, M184VI, Y181C, and G190A) compared to Sanger sequencing and next-generation sequencing (NGS).
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Dolutegravir (DTG) use in combination with tenofovir and lamivudine (TLD) is scaling up in Africa. However, HIV drug resistance (HIVDR) data to DTG remain scarce in Zimbabwe. We assessed the prevalence and genetic mechanisms of DTG... more
Dolutegravir (DTG) use in combination with tenofovir and lamivudine (TLD) is scaling up in Africa. However, HIV drug resistance (HIVDR) data to DTG remain scarce in Zimbabwe. We assessed the prevalence and genetic mechanisms of DTG resistance in people living with HIV initiating on TLD. A prospective cohort study was conducted between October 2021 and April 2023 among antiretroviral therapy (ART) naïve adults (≥18 years) attending care at an HIV clinic in Zimbabwe. Pre-treatment drug resistance (PDR) was assessed prior to TLD initiation and viral load (VL) outcome and acquired drug resistance (ADR) to TLD were described after 24 weeks follow-up. In total, 172 participants were enrolled in the study. The median (IQR) age and log10 VL were 39 (29–48) years and 5.41 (4.80–5.74) copies/mL, respectively. At baseline, no PDR to DTG was found. However, as previously reported, PDR to non-nucleotide reverse transcriptase inhibitor (NNRTI) was high (15%) whilst PDR to NRTI was low (4%). After...
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Background In Zimbabwe, children, adolescents and young adults living with HIV (CALWH) who are on public health antiretroviral therapy (ART) have inadequate viral load (VL) suppression. We assessed whether a clinic-based VL monitoring... more
Background In Zimbabwe, children, adolescents and young adults living with HIV (CALWH) who are on public health antiretroviral therapy (ART) have inadequate viral load (VL) suppression. We assessed whether a clinic-based VL monitoring could decrease 12-month virologic failure rates among these CALWH. Methods The study was registered on ClinicalTrials.gov: NCT03986099. CALWH in care at Chidamoyo Christian Hospital (CCH) and 8 rural outreach sites (ROS) on long-term community-based ART were randomized (1:1) to 6 monthly VL monitoring by COBAS®Ampliprep®/Taqman48® HIV-1 at the provincial referral laboratory (PRL) as per standard of care (SOC) or by the clinic-based SAMBA II assay, Diagnostics for the Real World, at CCH. VL suppression, turn-around-time (TAT) for VL results, drug switching and drug resistance in second-line failure were assessed at 12 months. Results Of 390 CALWH enrolled 347 (89%) completed 12 months follow-up. Median (IQR) age and ART duration were 14.1 (9.7–18.2) and...
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Introduction Maintaining virologic suppression of children and adolescents on ART in rural communities in sub-Saharan Africa is challenging. We explored switching drug regimens to protease inhibitor (PI) based treatment and reducing... more
Introduction Maintaining virologic suppression of children and adolescents on ART in rural communities in sub-Saharan Africa is challenging. We explored switching drug regimens to protease inhibitor (PI) based treatment and reducing nevirapine and zidovudine use in a differentiated community service delivery model in rural Zimbabwe. Methods From 2016 through 2018, we followed 306 children and adolescents on ART in Hurungwe, Zimbabwe at Chidamoyo Christian Hospital, which provides compact ART regimens at 8 dispersed rural community outreach sites. Viral load testing was performed (2016) by Roche and at follow-up (2018) by a point of care viral load assay. Virologic failure was defined as viral load ≥1,000 copies/ml. A logistic regression model which included demographics, treatment regimens and caregiver’s characteristics was used to assess risks for virologic failure and loss to follow-up (LTFU). Results At baseline in 2016, 296 of 306 children and adolescents (97%) were on first-li...
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Introduction pre-treatment drug resistance (PDR) can compromise the 3rd 95-95-95 global target for viral load suppression. The high complexity and cost of genotyping assays limits routine testing in many resource limited settings (RLS).... more
Introduction pre-treatment drug resistance (PDR) can compromise the 3rd 95-95-95 global target for viral load suppression. The high complexity and cost of genotyping assays limits routine testing in many resource limited settings (RLS). We assessed the performance of a rapid HIV-1 drug resistance assay, the Pan Degenerate Amplification and Adaptation (PANDAA) assay when screening for significant HIV-1 drug resistance mutations (DRMs) such as K65R, K103NS, M184VI, Y181C and G190A. Methods: we used previously generated amplicons from a cross-sectional study conducted between October 2018 and February 2020 of HIV-1 infected antiretroviral therapy (ART)-naïve or those reinitiating 1st line ART (18 years or older). The performance of the PANDAA assay in screening K65R, K103NS, M184VI, Y181C, and G190A mutations compared to the reference assay, Sanger sequencing was evaluated by Cohen´s kappa coefficient on Stata version 14 (StataCorp LP, College Station, TX, USA). Results one hundred and...
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Introduction: Following the World Health Organization (WHO)’s 3 by 5 initiative, antiretroviral treatment (ART) for HIV disease management has been scaled up rapidly in resource limited settings such as Zimbabwe. First line treatment... more
Introduction: Following the World Health Organization (WHO)’s 3 by 5 initiative, antiretroviral treatment (ART) for HIV disease management has been scaled up rapidly in resource limited settings such as Zimbabwe. First line treatment options include a three antiretroviral (ARV) drug regimens compromising of two nucleoside reverse transcriptase inhibitors (NRTIs) and one non-nucleoside (NNRTI). There are fears that
Background Healthcare workers are disproportionately affected by COVID-19. In low- and middle- income countries, they may be particularly impacted by underfunded health systems, lack of personal protective equipment, challenging working... more
Background Healthcare workers are disproportionately affected by COVID-19. In low- and middle- income countries, they may be particularly impacted by underfunded health systems, lack of personal protective equipment, challenging working conditions and barriers in accessing personal healthcare. Methods In this cross-sectional study, occupational health screening was implemented at the largest public sector medical centre in Harare, Zimbabwe, during the “first wave” of the country’s COVID-19 epidemic. Clients were voluntarily screened for symptoms of COVID-19, and if present, offered a SARS-CoV-2 nucleic acid detection assay. In addition, measurement of height, weight, blood pressure and HbA1c, HIV and TB testing, and mental health screening using the Shona Symptom Questionnaire (SSQ-14) were offered. An interviewer-administered questionnaire ascertained client knowledge and experiences related to COVID-19. Results Between 27th July and 30th October 2020, 951 healthcare workers access...
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BACKGROUND Previously used as part of salvage therapy, integrase strand transfer inhibitors (INSTIs) have become part of the preferred antiretroviral therapy (ART) first-line regimen in most low-to-middle income countries. With the... more
BACKGROUND Previously used as part of salvage therapy, integrase strand transfer inhibitors (INSTIs) have become part of the preferred antiretroviral therapy (ART) first-line regimen in most low-to-middle income countries. With the extensive use of dolutegravir in first-line ART, drug resistance mutations to INSTIs are inevitable. Therefore, active monitoring and surveillance of INSTI drug resistance is required. The aim of this study was to evaluate the genetic diversity of the integrase gene and determine pre-treatment INSTI resistance in Harare, Zimbabwe. METHODS Forty-four (44) HIV-1 Integrase sequences from 65 were obtained from treatment naive individuals using a custom genotyping method. Drug resistance mutations were determined using the Stanford HIV Drug Resistance Interpretation program. Viral subtyping was done by phylogenetic analysis and the REGA HIV subtyping tool determined recombinants. Natural polymorphisms were evaluated relative to the global subtype B and C consensus sequences. One hundred and ninety-two sequences from the region were accessed from GenBank to assess differences between the Zimbabwean sequences and those from neighbouring countries. RESULTS No major INSTI resistance mutations were detected, however, the L74I polymorphism was detected in three sequences of the 44 (6.8%). There was little genetic variability in the Integrase gene, with a mean genetic distance range of 0.053015. The subtype C consensus was identical to the global subtype C consensus and varied from the global subtype B consensus at five major positions; T124A, V201I, T218I, D278A and S283G. CONCLUSION The present study has provided baseline sequence data on the presence of HIV-1 subtype C Integrase gene drug resistance mutations from Harare, Zimbabwe.
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BACKGROUND Pre-treatment HIV drug resistance (PDR) can compromise antiretroviral therapy (ART) efficacy and undermine the WHO targets to end the AIDS epidemic as a public health threat by 2030. Thus, we examined the level of PDR in... more
BACKGROUND Pre-treatment HIV drug resistance (PDR) can compromise antiretroviral therapy (ART) efficacy and undermine the WHO targets to end the AIDS epidemic as a public health threat by 2030. Thus, we examined the level of PDR in Harare, Zimbabwe. METHODS Eligible study participants were adults who were ART-naïve or individuals with previous ART exposure reinitiating treatment, recruited between October 2018 and February 2020 in a HIV ART treatment clinic, in Harare. HIV drug resistance (HIVDR) tests were performed for all specimens with viral load >= 400 copies/ml and interpreted using the Stanford HIVDB Algorithm. Chi-square test or Fisher's exact test was used for comparison of proportions of PDR across ART- naïve or prior ART exposed participants. All statistical analyses were performed using Stata version 14. RESULTS Overall, 120 samples were genotyped of whom 104 were ART-naïve and 16 reported previous ART exposure. The overall PDR frequency among all participants was 31% (95%CI: 22.5-39.6). PDR to any NNRTI was reported in 29% (95%CI: 21.0-37.9). PDR to NRTIs and PIs were low, found in 3% (95%CI: 0.9-8.2) and 1% (95%CI: 0.02-4.52) respectively. PDR to NNRTIs (EFV/NVP) was found in 17% (95%CI: 10.5-24.6) and was more than 6 times higher among people with previous ART exposure than ART- naïve people: 63% versus 10%, p<0.001. CONCLUSION Our study shows that PDR to NNRTIs in Zimbabwe has remarkably increased from the 10.9% prevalence reported in the 2016 WHO survey. Addressing PDR at a national level is a critical need and will be facilitated by fast-tracking the transition to dolutegravir in 1st line ART regimens.
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BackgroundEfavirenz (EFV)-based regimens select broad drug resistance to nonnucleoside reverse-transcriptase inhibitors (NNRTIs), limiting the effectiveness of EFV and other NNRTIs. The duration, persistence, and decay of drug resistance... more
BackgroundEfavirenz (EFV)-based regimens select broad drug resistance to nonnucleoside reverse-transcriptase inhibitors (NNRTIs), limiting the effectiveness of EFV and other NNRTIs. The duration, persistence, and decay of drug resistance mutations (DRMs) in the proviral reservoir is not well defined.MethodsParticipants with virologic failure of EFV-based regimens and drug-resistant viremia with the K103N mutation in plasma ribonucleic acid (RNA) were identified from AIDS Clinical Trials Group (ACTG) studies A364 and A5095. These individuals received a second-line, boosted protease inhibitor-based regimen with suppression of viremia for up to10 years during long-term follow-up (median = 3.6 years; interquartile range, 2.1–6.9 years). Proviral deoxyribonucleic acid (DNA) from cryopreserved peripheral blood mononuclear cells was sequenced to identify the persistence of DRM.ResultsTwenty-eight participants from ACTG 364 and ACTG 5095 were evaluated. Sanger sequencing of proviral DNA det...
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Treatment of HIV has reduced HIV/AIDS-related mortality. Sustaining >90% virologic suppression in sub-Saharan Africa requires decentralized care and prevention services to rural communities. In Zimbabwe, the number of people receiving... more
Treatment of HIV has reduced HIV/AIDS-related mortality. Sustaining >90% virologic suppression in sub-Saharan Africa requires decentralized care and prevention services to rural communities. In Zimbabwe, the number of people receiving antiretroviral treatment (ART) has increased rapidly. However, access to treatment monitoring tools such as viral load and drug resistance testing is limited. We assessed virologic treatment outcomes among ART recipients in Nyamutora, a rural community receiving bimonthly ART and prevention services. We enrolled all ART recipients (143) at 6-monthly visits in the Nyamutora community in 2014 and 2015. Whole blood samples were collected in K-EDTA tubes, transported to Harare for CD4 counts and viral load testing, and genotype was obtained in participants with viral loads >1,000 copies/ml. Ages ranged from 2 to 75 years (median 43 years) with a median 42 months on ART at follow-up. Eight of 143 (6%) had viral loads >1,000 copies/ml at one of the ...
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To characterize HIV-1 transmission dynamics in regions where the burden of HIV-1 is greatest, the 'Phylogenetics and Networks for Generalised HIV Epidemics in Africa' consortium (PANGEA-HIV) is sequencing full-genome viral... more
To characterize HIV-1 transmission dynamics in regions where the burden of HIV-1 is greatest, the 'Phylogenetics and Networks for Generalised HIV Epidemics in Africa' consortium (PANGEA-HIV) is sequencing full-genome viral isolates from across sub-Saharan Africa. We report the first 3,985 PANGEA-HIV consensus sequences from four cohort sites (Rakai Community Cohort Study, n=2,833; MRC/UVRI Uganda, n=701; Mochudi Prevention Project, n=359; Africa Health Research Institute Resistance Cohort, n=92). Next-generation sequencing success rates varied: more than 80% of the viral genome from the gag to the nef genes could be determined for all sequences from South Africa, 75% of sequences from Mochudi, 60% of sequences from MRC/UVRI Uganda, and 22% of sequences from Rakai. Partial sequencing failure was primarily associated with low viral load, increased for amplicons closer to the 3' end of the genome, was not associated with subtype diversity except HIV-1 subtype D, and remaine...
Research Interests: Immunology, Phylogenetics, Biology, Risk, Infectious Diseases, and 15 moreMedicine, Phylogenomics, Phylodynamics, Dna Sequencing, Reconstruction, Infectious Disease, Fishing Communities, Genome, Clinical Sciences, Human immunodeficiency virus, Lake Victoria, Science Technology, Pangea, Amplicon, and Incongruence
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Research Interests: Genetics, Microbiology, Medical Microbiology, Biology, Virology, and 15 moreMedicine, HIV, Zimbabwe, Humans, HIV drug resistance, United States, Drug Resistance, Journal of Virological Methods, Genotype, Highly Active Antiretroviral Therapy, Genotyping, Microbial Sensitivity Tests, Treatment Failure, Anti-retroviral agents, and HIV infections
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Research Interests: Virology, Medicine, HIV, South Africa, Population, and 15 moreHumans, Mutation, HIV drug resistance, Female, Drug Resistance, Male, Clinical Sciences, Genotype, Adult, Public Health Surveillance, Highly Active Antiretroviral Therapy, Retrospective Studies, Viral Load, reverse transcriptase inhibitors, and HIV infections
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Research Interests: Biochemistry, Bioinformatics, Evolutionary Biology, Environmental Science, Developmental Biology, and 15 moreClimate Change, Computational Biology, Biotechnology, Cancer, Biology, Cell Cycle, Ecology, Drug Discovery, Evolution, Astrophysics, Astronomy, DNA, Drug Resistance, Cell Signalling, and Earth Science
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South Africa has the highest number of individuals with HIV-1 infection worldwide with the epicentre of the epidemic in the province of KwaZulu-Natal (KZN). Currently South Africa has the largest antiretroviral treatment (ART) programme... more
South Africa has the highest number of individuals with HIV-1 infection worldwide with the epicentre of the epidemic in the province of KwaZulu-Natal (KZN). Currently South Africa has the largest antiretroviral treatment (ART) programme with KZN and Gauteng provinces accounting for 56% of all patients on ART. In order to understand transmission dynamics and temporal trends in drug resistance, we performed a comprehensive literature review of primary and acquired drug resistance in adults and children in KZN over a ten year period, 2003-2013.
Transmitted drug resistance (TDR) to non-nucleoside reverse transcriptase inhibitors (NNRTIs) in adults is increasing to moderate levels (5-15%). Acquired drug resistance to first line drug regimens in adults was associated with at least one mutation to nucleoside reverse transcriptase inhibitors (NRTI) or NNRTIs or both, with M184V and K103N the most common mutations. Prevalence of thymidine analogues (TAMs) was low to moderate. There was no published data on acquired drug resistance to second line agents. Children failing first line drug regimens harboured drug resistance mutations to NRTI (M184V and TAMs), NNRTI (V106M and K103N) and protease inhibitors (PIs) (V82A).
Concerns have been raised about the high levels of the K65R mutation associated with tenofovir regimens in adults and children. Continued vigilance is needed for ART failure in both adults and children, so as not to compromise second line treatment options. Drug resistance genotyping is now cost effective and should be considered for inclusion in regional surveillance studies and during routine care.
Transmitted drug resistance (TDR) to non-nucleoside reverse transcriptase inhibitors (NNRTIs) in adults is increasing to moderate levels (5-15%). Acquired drug resistance to first line drug regimens in adults was associated with at least one mutation to nucleoside reverse transcriptase inhibitors (NRTI) or NNRTIs or both, with M184V and K103N the most common mutations. Prevalence of thymidine analogues (TAMs) was low to moderate. There was no published data on acquired drug resistance to second line agents. Children failing first line drug regimens harboured drug resistance mutations to NRTI (M184V and TAMs), NNRTI (V106M and K103N) and protease inhibitors (PIs) (V82A).
Concerns have been raised about the high levels of the K65R mutation associated with tenofovir regimens in adults and children. Continued vigilance is needed for ART failure in both adults and children, so as not to compromise second line treatment options. Drug resistance genotyping is now cost effective and should be considered for inclusion in regional surveillance studies and during routine care.
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Human immunodeficiency virus 2 (HIV-2) is found predominantly in West Africa. It is not unlikely, however, that HIV-2 may also be found in South Africa, due to the influx of immigrants into this country. It is important to distinguish... more
Human immunodeficiency virus 2 (HIV-2) is found predominantly in West Africa. It is not unlikely, however, that HIV-2 may also be found in South Africa, due to the influx of immigrants into this country. It is important to distinguish between HIV-1 and HIV-2 since the clinical courses and treatment responses of these viruses are different. Routine serological methods for diagnosing HIV do not differentiate between HIV-1 and -2 infections, while rapid tests, viral load quantification and PCR are HIV-type—specific. The objective of this study was to describe the seroprevalence and molecular epidemiology of HIV-2 in KwaZulu-Natal, one of the regions with the highest HIV prevalence in the world and home of the two largest harbors in South Africa. HIV-1 positive samples were screened for antibodies against HIV-2, using a rapid test. The confirmation of HIV-2 positive samples was done by PCR. Of the 2,123 samples screened, 319 (15%) were identified as positive by the rapid test. None of these samples were confirmed positive by PCR. To explore this discrepancy in the results, a subset (n = 52) of the rapid HIV-2 positive samples was subjected to Western blotting. Thirty-seven (71%) of these were positive, yielding an overall HIV-2 seroprevalence of 10.6%. Three out of 28 (10.7%) Western blot positive samples were positive by a Pepti-LAV assay. This discrepancy between serological and molecular confirmation may be attributed to non-specific or cross-reacting antibodies. The use of rapid tests and Western blots for HIV-2 diagnosis in South Africa should be interpreted with caution. J. Med. Virol. 85:2065–2071, 2013. © 2013 Wiley Periodicals, Inc.Human immunodeficiency virus 2 (HIV-2) is found predominantly in West Africa. It is not unlikely, however, that HIV-2 may also be found in South Africa, due to the influx of immigrants into this country. It is important to distinguish between HIV-1 and HIV-2 since the clinical courses and treatment responses of these viruses are different. Routine serological methods for diagnosing HIV do not differentiate between HIV-1 and -2 infections, while rapid tests, viral load quantification and PCR are HIV-type—specific. The objective of this study was to describe the seroprevalence and molecular epidemiology of HIV-2 in KwaZulu-Natal, one of the regions with the highest HIV prevalence in the world and home of the two largest harbors in South Africa. HIV-1 positive samples were screened for antibodies against HIV-2, using a rapid test. The confirmation of HIV-2 positive samples was done by PCR. Of the 2,123 samples screened, 319 (15%) were identified as positive by the rapid test. None of these samples were confirmed positive by PCR. To explore this discrepancy in the results, a subset (n = 52) of the rapid HIV-2 positive samples was subjected to Western blotting. Thirty-seven (71%) of these were positive, yielding an overall HIV-2 seroprevalence of 10.6%. Three out of 28 (10.7%) Western blot positive samples were positive by a Pepti-LAV assay. This discrepancy between serological and molecular confirmation may be attributed to non-specific or cross-reacting antibodies. The use of rapid tests and Western blots for HIV-2 diagnosis in South Africa should be interpreted with caution. J. Med. Virol. 85:2065–2071, 2013. © 2013 Wiley Periodicals, Inc.