Cardiac remodeling has been reported in HIV-exposed but uninfected (HEU) infants. Inflammatory biomarkers have also been observed to be associated with cardiac remodeling in older HEU children but have not been widely studied in infants. We identified left ventricular (LV) cardiac changes in HEU infants exposed to current ARV therapy in utero and determined their association with the inflammatory biomarkers, C-reactive protein (CRP), and cardiac troponin. This comparative cross-sectional study included 196 HEU infants and 198 matched HIV-unexposed controls. We evaluated LV structure and function using echocardiography and tested blood samples using highly sensitive CRP (hsCRP) and cardiac troponin I (cTnI-plus) measurements. Data were analyzed using the R statistical software package; p values of < 0.05 were considered statistically significant. The HEU infants had thicker LV posterior walls and interventricular septa in diastole (median difference 0.55 and 0.30, 95

BackgroundHIV-related stigma is often expressed as irrational behaviors, negative attitudes, and unfavorable judgments toward people living with or at risk of HIV which remains very common in low- and middle-income countries including Nigeria. This study assessed the level of HIV-related stigma and its associated factors among Young Men who have Sex with Men (YMSM) in HIV care.MethodologyThis was a cross-sectional study conducted among 122 YMSM to assess the level of HIV-related stigma and its associated factors among YMSM in HIV care using respondent driven sampling between July 2023 and April 2024. Quantitative method of data collection was employed and SPSS version 23 was used for data analysis. A p-value of ≤ 0.05 was considered statistically significant.ResultsThe mean age of the study participants was 22.2 ± 2.0 years with 56 (45.6%) being 22 years and below. The total HIV- related stigma score for the participants was 121.9 ± 18.8 with high-level of stigma reported among 40 (32.8%) of the participants. Significant variation in the mean total HIV-related stigma score was found with duration on HIV treatment with those who had been on treatment for >3 years having a mean score of 117.8 ± 15.2 compared to 110.3 ± 20.1 for those who had been on treatment for <2 years (mean difference: −7.50; 95% Confidence interval: −14.45, −5.51; p = 0.035).ConclusionThis study found a high level of HIV-related stigma among the YMSM in HIV care which is significantly influenced by the duration of time in HIV care.

Yellow fever virus (YFV) and West Nile virus (WNV) co-circulate with other arboviruses, including Zika (ZIKV), dengue (DENV), and chikungunya virus (CHIKV), in sub-Saharan Africa. Associations between preexisting YFV and WNV immunity with symptoms and adverse infant outcomes among pregnant women exposed to orthoflaviviruses are unknown. We retrospectively studied a prospective cohort of pregnant women enrolled between 2019 and 2022 in Jos, Nigeria. Rapid tests identified ZIKV, DENV, and CHIKV IgM/IgG reactivity for enrolment; 216 women underwent Western blot for YFV and WNV IgG. Logistic regression evaluated associations between arboviral seropositivity and maternal symptoms or adverse infant outcomes. Sequential serology of mother-infant pairs estimated the persistence of passively transferred maternal YFV antibodies. YFV IgG was detected in 50.5% (109/216) and WNV IgG in 5.1% (11/216) of maternal samples. YFV and WNV seropositivity was significantly associated with maternal symptoms (OR = 2.02, 95% CI: 1.35-3.02, P = 0.001), as was YFV seropositivity alone (OR = 1.77, 95% CI: 1.21-2.61, P < 0.004). CHIKV IgM reactivity was significantly associated with abnormal infant outcomes (OR = 2.38, 95% CI: 1.43-4.02, p = 0.001), but not ZIKV and DENV IgM reactivity. Passive maternal YFV IgG waned in infants at a median of 3.1 months (IQR: 1.65-5.35 months) after birth. YFV and WNV seropositivity was associated with maternal symptoms but not with adverse infant outcomes. Rapid waning of maternal YFV IgG highlights infant vulnerability and supports enhanced surveillance and maternal immunization strategies.

BackgroundThe COVID-19 pandemic had a severe impact globally, yet African populations exhibited unexpectedly lower rates of severe disease and mortality. We investigated the potential role of pre-existing immunity in shaping the epidemiology of COVID-19 in Africa.MethodsPlasma collected from Senegalese female sex workers prior to the COVID-19 pandemic was screened for SARS-CoV-2 and human coronavirus (hCoV) antibodies by virion immunoblots. For antibody-reactive plasma, paired peripheral blood mononuclear cells were stimulated by fusion proteins and IFN-γ cellular responses were assessed via ELISPOT.ResultsWe observed substantial levels of pre-existing cross-reactive immunity to SARS-CoV-2, stemming from prior exposure to seasonal hCoVs. Our antibody analysis revealed a 23.5% (47/200) seroprevalence rate against SARS-CoV-2 nucleocapsid (N). These samples were then probed for antibodies against hCoV spike (S) and/or N antigens; 85.1% (40/47), 70.2% (33/47), and 95.7% (45/47) were antibody reactive against hCoV-229E, hCoV-OC43, or hCoV-HKU1, respectively. Our analysis of cellular responses also demonstrated cross-reactivity to SARS-CoV-2 with 80.0% (36/45) and 82.2% (37/45) showing IFN-γ responses against S and N, respectively. A unique pre-pandemic subject had cross-reactive SARS-CoV-2 S antibodies with detectable neutralization and cross-reactive cellular responses.ConclusionThese findings suggest that prior hCoV exposure may induce cross-reactive adaptive immunity, potentially contributing to protection against COVID-19. Our study provides unique data on the dynamics of hCoV and SARS-CoV-2 immunity in Senegal and underscores the importance of understanding the role of pre-existing immunity in shaping COVID-19 outcomes globally.

The COVID-19 pandemic had a severe impact globally, yet African populations exhibited unexpectedly lower rates of severe disease and mortality. We investigated the potential role of pre-existing immunity in shaping the epidemiology of COVID-19 in Africa. Leveraging paired plasma and peripheral blood mononuclear cells collected from Senegalese female sex workers prior to the COVID-19 pandemic, we observed substantial levels of pre-existing cross-reactive immunity to SARS-CoV-2, stemming from prior exposure to seasonal human coronaviruses (hCoVs). Our antibody analysis revealed a 23.5% (47/200) seroprevalence rate against SARS-CoV-2 nucleocapsid (N). Of these SARS-CoV-2 N-reactives, 85.1% (40/47), 44.7% (21/47), and 95.7% (45/47) showed antibody reactivity against hCoV-229E or hCoV-OC43 spike (S) and/or N or hCoV-HKU1 S. Our analysis of cellular responses also demonstrated cross-reactivity to SARS-CoV-2 with 82.2% (37/45) and 84.4% (38/45) showing IFN-γ responses against S and N, respectively. These findings suggest that prior hCoV exposure may induce cross-reactive adaptive immunity, potentially contributing to protection against COVID-19. A unique pre-pandemic subject had cross-reactive SARS-CoV-2 S antibodies with detectable neutralization and cellular responses. Our study provides unique insights into the dynamics of hCoV and SARS-CoV-2 immunity in West African populations and underscores the importance of understanding the role of pre-existing immunity in shaping COVID-19 outcomes globally. ### Competing Interest Statement The authors have declared no competing interest. ### Funding Statement This study was funded by a grant to Harvard University, Center for African Studies, Motsepe Presidential Research Accelerator Fund for Africa ### Author Declarations I confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained. Yes The details of the IRB/oversight body that provided approval or exemption for the research described are given below: At the time of the original study, all women provided informed consent for the collection and archiving of samples and corresponding data; ethical clearance from the Harvard Institutional Review Board (IRB) and the research ethics committee at Chiekh Anta Diop University, Dakar, Senegal were obtained. All archived samples and corresponding data were anonymized, and as such this secondary analysis did not constitute human subject research by institutional guidelines and regulations which follow the Federal Policy for Protection of Human Subjects. I confirm that all necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived, and that any patient/participant/sample identifiers included were not known to anyone (e.g., hospital staff, patients or participants themselves) outside the research group so cannot be used to identify individuals. Yes I understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance). Yes I have followed all appropriate research reporting guidelines, such as any relevant EQUATOR Network research reporting checklist(s) and other pertinent material, if applicable. Yes All data produced in the present work are available upon reasonable request to the authors.

We determined pretreatment and acquired human immunodeficiency virus (HIV) drug resistance among children with HIV type 1 (HIV-1) in Jos, Nigeria. The majority (71%) of those who failed first-line antiretroviral therapy were on a nevirapine-containing regimen. The prevalence of pretreatment (48%) and acquired (76%) HIV drug resistance mutations was high in our study. Wider access to HIV drug resistance testing after treatment failure is necessary to optimize second-line treatment options among children with HIV in Nigeria.

Objectives: Chikungunya virus (CHIKV), a reemerging global public health concern, which causes acute febrile illness, rash, and arthralgia and may affect both mothers and infants during pregnancy. Mother-to-child transmission (MTCT) of CHIKV in Africa remains understudied. Methods: Our cohort study screened 1006 pregnant women with a Zika/dengue/CHIKV rapid test at two clinics in Nigeria between 2019 and 2022. Women who tested positive for the rapid test were followed through their pregnancy and their infants were observed for 6 months, with a subset tested by reverse transcription-polymerase chain reaction (RT-PCR) and neutralization, to investigate seropositivity rates and MTCT of CHIKV. Results: Of the 1006, 119 tested positive for CHIKV immunoglobulin (Ig)M, of which 36 underwent detailed laboratory tests. While none of the IgM reactive samples were RT-PCR positive, 14 symptomatic pregnant women were confirmed by CHIKV neutralization test. Twelve babies were followed with eight normal and four abnormal outcomes, including stillbirth, cleft lip/palate with microcephaly, preterm delivery, polydactyly with sepsis, and jaundice. CHIKV IgM testing identified three possible antepartum transmissions. Conclusion: In Nigeria, we found significant CHIKV infection in pregnancy and possible CHIKV antepartum transmission associated with birth abnormalities.

We determined pretreatment and acquired human immunodeficiency virus (HIV) drug resistance among children with HIV type 1 (HIV-1) in Jos, Nigeria. The majority (71%) of those who failed first-line antiretroviral therapy were on a nevirapine-containing regimen. The prevalence of pretreatment (48%) and acquired (76%) HIV drug resistance mutations was high in our study. Wider access to HIV drug resistance testing after treatment failure is necessary to optimize second-line treatment options among children with HIV in Nigeria.

Chikungunya virus (CHIKV) has become a global public health concern since the reemergence of the Indian Ocean lineage and expansion of the Asian genotype. CHIKV infection causes acute febrile illness, rash, and arthralgia and during pregnancy may affect both mothers and infants. The mother-to-child transmission (MTCT) of CHIKV in Africa remains understudied. We screened 1006 pregnant women at two clinics in Nigeria between 2019 and 2022 and investigated the prevalence and MTCT of CHIKV. Of the 1006, 119 tested positive for CHIKV IgM, of which 36 underwent detailed laboratory tests. While none of the IgM reactive samples were RT-PCR positive, 14 symptomatic pregnant women were confirmed by CHIKV neutralization test. Twelve babies were followed with 8 normal and 4 abnormal outcomes, including stillbirth, cleft lip/palate with microcephaly, preterm delivery, polydactyly with sepsis and jaundice. CHIKV IgM testing identified 3 antepartum transmissions, further studies will determine its impact in antepartum infection.

The adverse impact of Zika (ZIKV), dengue (DENV), and chikungunya (CHIKV) virus infection in pregnancy has been recognized in Latin America and Asia but is not well studied in Africa. In Nigeria, we screened 1006 pregnant women for ZIKV, DENV and CHIKV IgM/IgG by rapid test (2019-2022). Women with acute infection were recruited for prospective study and infants were examined for any abnormalities from delivery through six months. A subset of rapid test-reactive samples were confirmed using virus-specific ELISAs and neutralization assays. Prevalence of acute infection (IgM+) was 3.8%, 9.9% and 11.8% for ZIKV, DENV and CHIKV, respectively; co-infections represented 24.5% of all infections. Prevalence in asymptomatic women was twice the level of symptomatic infection. We found a significant association between acute maternal ZIKV/DENV/CHIKV infection and any gross abnormal birth outcome (p=0.014). Further prospective studies will contribute to our understanding of the clinical significance of these endemic arboviruses in Africa.

BACKGROUND:Human immunodeficiency virus (HIV) viral load (VL) monitoring is critical for antiretroviral therapy (ART) management. Point-of-care (POC) VL testing has been reported to be feasible and preferred over standard-of-care (SOC) testing in many low- and middle-income country settings where rapid results could improve patient outcomes. METHODS:The timeliness of receipt of VL results was evaluated in an open-label, randomized, controlled trial among patients newly initiating ART. Clinical outcomes with POC VL monitoring using Cepheid Xpert vs SOC VL at Jos University Teaching Hospital and Comprehensive Health Centre Zamko in Nigeria were assessed. We determined time between specimen collection and recording of VL in patient charts, receipt of results, and ART switch for those who met virologic failure criteria. RESULTS:Between April 2018 and October 2019, we screened 696 ART-naive individuals; 273 were randomized to POC and 268 to SOC HIV-1 VL testing. Participants in the POC arm received VL results significantly faster than those in the SOC arm (0.1 median days, interquartile range [IQR], 0.1-0.2 vs 143.1 days, IQR, 56.0-177.1, respectively; P < .0001). Participants in the POC arm with confirmed virologic failure vs those in the SOC arm were switched more rapidly to a second-line regimen (0 median days, IQR, 0-28 vs 66 days, IQR, 63-123, respectively; P = .03). CONCLUSIONS:POC VL testing resulted in significant improvement in the timeliness of VL result receipt by patients and use for effective HIV clinical management. In patients experiencing VL failure, POC monitoring enabled prompt switching to second-line ART regimens. CLINICAL TRIALS REGISTRATION:NCT03533868.

Background: Early evidence suggested that the impact of the COVID-19 pandemic was less severe in Africa compared to other parts of the world. However, more recent studies indicate higher SARS-CoV-2 infection and COVID-19 mortality rates on the continent than previously documented. Research is needed to better understand SARS-CoV-2 infection and immunity in Africa. Methods: In early 2021, we studied the immune responses in healthcare workers (HCWs) at Lagos University Teaching Hospital ( n = 134) and Oxford-AstraZeneca COVID-19 vaccine recipients from the general population ( n = 116) across five local government areas (LGAs) in Lagos State, Nigeria. Western blots were used to simultaneously detect SARS-CoV-2 spike and nucleocapsid (N) antibodies ( n = 250), and stimulation of peripheral blood mononuclear cells with N followed by an IFN-������ ELISA was used to examine T cell responses ( n = 114). Results: Antibody data demonstrated high SARS-CoV-2 seroprevalence of 72 & BULL;4% (97/134) in HCWs and 60 & BULL;3% (70/116) in the general population. Antibodies directed to only SARS-CoV-2 N, suggesting pre-existing coronavirus immunity, were seen in 9 & BULL;7% (13/134) of HCWs and 15 & BULL;5% (18/116) of the general population. T cell responses against SARS-CoV-2 N ( n = 114) were robust in detecting exposure to the virus, demonstrating 87 & BULL;5% sensitivity and 92 & BULL;9% specificity in a subset of control samples tested. T cell responses against SARS-CoV-2 N were also observed in 83.3% of individuals with N-only antibodies, further suggesting that prior non-SARS-CoV2 coronavirus infection may provide cellular immunity to SARS-CoV-2. Conclusions: These results have important implications for understanding the paradoxically high SARS-CoV-2 infection with low mortality rate in Africa and supports the need to better understand the implications of SARSCoV-2 cellular immunity.

Background. Point-of-care (POC) viral load (VL) tests provide results within hours, enabling same-day treatment interventions. We assessed treatment outcomes with POC vs standard-of-care (SOC) VL monitoring. Methods. We implemented a randomized controlled trial at an urban and rural hospital in Nigeria. Participants initiating antiretroviral therapy (ART) were randomized 1:1 for monitoring via the POC Cepheid Xpert or SOC Roche COBAS (v2.0) HIV-1 VL assays. Viral suppression (VS) and retention in care at 12 months were compared via intention-to-treat (ITT) and per-protocol (PP) analyses. Post-trial surveys for POC patients and healthcare workers (HCWs) evaluated acceptability. Results. During April 2018-October 2019, 268 SOC and 273 POC patients enrolled in the trial. Viral suppression at <1000 copies/mL at 12 months was 59.3% (162/273) for POC and 52.2% (140/268) for SOC (P = .096) in ITT analysis and 77.1% (158/205) for POC and 65.9% (137/208) for SOC (P = .012) in PP analysis. Retention was not significantly different in ITT analysis but was 85.9% for POC and 76.9% for SOC (P = .02) in PP analysis. The increased VS in the POC arm was attributable to improved retention and documentation of VL results. POC monitoring was preferred over SOC by 90.2% (147/163) of patients and 100% (15/15) of HCWs thought it facilitated patient care. Conclusions. POC VL monitoring did not improve 12-month VS among those with results but did improve retention and VS documentation and was preferred by most patients and HCWs. Further research can inform best POC implementation conditions and approaches to optimize patient care.

Background The HIV-1 epidemic in sub-Saharan Africa is heterogeneous with diverse unevenly distributed subtypes and regional differences in prevalence. Subtype-specific differences in disease progression rate and transmission efficiency have been reported, but the underlying biological mechanisms have not been fully characterized. Here, we tested the hypothesis that the subtypes prevalent in the East Africa, where adult prevalence rate is higher, have lower viral replication capacity (VRC) than their West African counterparts where adult prevalence rates are lower. Results Gag-protease sequencing was performed on 213 and 160 antiretroviral-naïve chronically infected participants from West and East Africa respectively and bioinformatic tools were used to infer subtypes and recombination patterns. VRC of patient-derived gag-protease chimeric viruses from West (n = 178) and East (n = 114) Africa were determined using a green fluorescent protein reporter-based cell assay. Subtype and regional differences in VRC and amino acid variants impacting VRC were identified by statistical methods. CRF02_AG (65%, n = 139), other recombinants (14%, n = 30) and pure subtypes (21%, n = 44) were identified in West Africa. Subtypes A1 (64%, n = 103), D (22%, n = 35), or recombinants (14%, n = 22) were identified in East Africa. Viruses from West Africa had significantly higher VRC compared to those from East Africa ( p < 0.0001), with subtype-specific differences found among strains within West and East Africa ( p < 0.0001). Recombination patterns showed a preference for subtypes D, G or J rather than subtype A in the p6 region of gag , with evidence that subtype-specific differences in this region impact VRC. Furthermore, the Gag A83V polymorphism was associated with reduced VRC in CRF02_AG. HLA-A*23:01 ( p = 0.0014) and HLA-C*07:01 ( p = 0.002) were associated with lower VRC in subtype A infected individuals from East Africa. Conclusions Although prevalent viruses from West Africa displayed higher VRC than those from East Africa consistent with the hypothesis that lower VRC is associated with higher population prevalence, the predominant CRF02_AG strain in West Africa displayed higher VRC than other prevalent strains suggesting that VRC alone does not explain population prevalence. The study identified viral and host genetic determinants of virus replication capacity for HIV-1 CRF02_AG and subtype A respectively, which may have relevance for vaccine strategies.

Nigeria has the highest number of AIDS-related deaths in the world. In this study, we characterised the HIV-1 molecular epidemiology by analysing 1442 HIV-1 pol sequences collected 1999–2014 from four geopolitical zones in Nigeria using state-of-the-art maximum-likelihood and Bayesian phylogenetic analyses. The main circulating forms were the circulating recombinant form (CRF) 02_AG (44% of the analysed sequences), CRF43_02G (16%), and subtype G (8%). Twenty-three percent of the sequences represented unique recombinant forms (URFs), whereof 37 (11%) could be grouped into seven potentially novel CRFs. Bayesian phylodynamic analysis suggested that five major Nigerian HIV-1 sub-epidemics were introduced in the 1960s and 1970s, close to the Nigerian Civil War. The analysis also indicated that the number of effective infections decreased in Nigeria after the introduction of free antiretroviral treatment in 2006. Finally, Bayesian phylogeographic analysis suggested gravity-like dynamics in which virus lineages first emerge and expand within large urban centers such as Abuja and Lagos, before migrating towards smaller rural areas. This study provides novel insight into the Nigerian HIV-1 epidemic and may have implications for future HIV-1 prevention strategies in Nigeria and other severely affected countries.

Background We identified a HIV-positive cohort in virologic failure (VF) who re-suppressed without drug switch. We characterized their drug resistance mutations (DRM) and adherence profiles to learn how to better manage HIV drug resistance. A retrospective cohort study utilizing clinical data and stored samples. Patients received ART at three Nigerian treatment centres. Plasma samples stored when they were in VF were genotyped. Result Of 126 patients with samples available, 57 were successfully genotyped. From ART initiation, the proportion of patients with adherence ≥90% increased steadily from 54% at first high viral load (VL) to 67% at confirmed VF, and 81% at time of re-suppressed VL. Sixteen (28%) patients had at least one DRM. Forty-six (81%) patients had full susceptibility to the three drugs in their first-line (1 L) regimen. Thirteen (23%) were resistant to at least one antiretroviral drug but three were resistant to drugs not used in Nigeria. Ten patients had resistance to their 1 L drug(s) and six were fully susceptible to the three drugs in the recommended second-line regimen. Conclusion This cohort had little drug resistance mutations. We conclude that if adherence is not assured, patients could exhibit virologic failure without having developed mutations associated with drug resistance.

Background: Many lines of evidence point to HIV-1 subtype-specific differences in the development of drug resistance mutations. While variation between subtype C and others has been extensively explored, there has been less emphasis on subtypes common to West Africa. We examined a previously described national survey of pretreatment drug resistance in HIV-1-infected Nigerian children aged <18 months, to explore the association between subtypes and patterns of resistance. Methods: Five hundred and forty-nine dried blood spots, from 15 early infant diagnostic facilities in Nigeria, were amplified and HIV-1 polymerase was sequenced. Four hundred and twenty-four were analysed for surveillance drug resistance mutations (SDRMs). Associations between subtype and SDRMs were evaluated by Fishers exact test and logistic regression analysis, controlling for geographical region and exposure. Results: Using the sub-subtypes of HIV-1 G defined by Delatorre et al. (PLoS One 2014; 9: e98908) the most common subtypes were CRF02_AG (174, 41.0%), G(WA-I) (128, 30.2%), G(WA-II) (24, 5.7%), G(CA) (11, 2.6%), A (21, 5.0%) and CRF06_cpx (18, 4.2%). One hundred and ninety infants (44.8%) had >= 1 NNRTI mutation, 92 infants (21.7%) had >= 1 NRTI mutation and 6 infants (1.4%) had >= 1 PI mutation. By logistic regression, 67N was more common in G(WA-II)/G(CA) than CRF02_AG (OR 12.0, P = 0.006), as was 70R (OR 23.1, P = 0.007), 184I/V (OR 2.92, P = 0.020), the presence of >= 1 thymidine analogue mutation (TAM) (OR 3.87, P = 0.014), >= 1 type 2 TAM (OR 7.61, P = 0.001) and >= 1 NRTI mutation (OR 3.26, P = 0.005). Conclusions: This dataset reveals differences among SDRMs by subtype; in particular, between the G(WA-II) and G(CA) subclades, compared with CRF02_AG and G(WA-I).

Abstract Nigeria has been reported to have the highest number of AIDS-related deaths in the world. In this study, we aimed to use molecular epidemiology to investigate the HIV-1 diversity and phylodynamics in Nigeria. We analyzed 1,442 HIV-1 pol sequences collected from 1999 to 2014 from seven geopolitical zones in Nigeria. The main circulating strains, CRF02_AG (44.1% of the analyzed sequences), Subtype G (8.3%), and CRF43_02G (16.4%), were introduced to Nigeria in the 1960s, 1970s, and 1980s, respectively. The number of effective infections decreased in Nigeria after the introduction of free antiretroviral treatment in 2006. We also found a significant number of unique recombinant forms (22.7%), the majority of which were recombinants between the two or three of the main circulating strains described above. In addition, phylogeographic analysis indicates multiple occasions of HIV-1 transmission between Lagos and Abuja (two of the main cities in Nigeria). Our results may be relevant for HIV-1 intervention and contribute in making informed decisions in strategies aiming at reducing further spread of HIV-1 in Nigeria.

BACKGROUND:ATCC HIV-1 drug resistance test kit was designed to detect HIV-1 drug resistance (HIVDR) mutations in the protease and reverse transcriptase genes for all HIV-1 group M subtypes and circulating recombinant forms. The test has been validated for both plasma and dried blood spot specimen types with viral load (VL) of ≥1000 copies/ml. We performed an in-country assessment on the kit to determine the genotyping sensitivity and its accuracy in detecting HIVDR mutations using plasma samples stored under suboptimal conditions.METHODS:Among 572 samples with VL ≥1000 copies/ml that had been genotyped by ViroSeq assay, 183 were randomly selected, including 85 successful genotyped and 98 unsuccessful genotyped samples. They were tested with ATCC kits following the manufacturer's instructions. Sequence identity and HIVDR patterns were analysed with Stanford University HIV Drug Resistance HIVdb program.RESULTS:Of the 183 samples, 127 (69.4%) were successfully genotyped by either method. While ViroSeq system genotyped 85/183 (46.5%) with median VL of 32,971 (IQR: 11,150-96,506) copies/ml, ATCC genotyped 115/183 (62.8%) samples with median VL of 23,068 (IQR: 7,397-86,086) copies/ml. Of the 98 unsuccessful genotyped samples with ViroSeq assay, 42 (42.9%) samples with lower median VL of 13,906 (IQR: 6,122-72,329) copies/ml were successfully genotyped using ATCC. Sequence identity analysis revealed that the sequences generated by both methods were >98% identical and yielded similar HIVDR profiles at individual patient level.CONCLUSION:This study confirms that ATCC kit showed greater sensitivity in genotyping plasma samples stored in suboptimal conditions experiencing frequent and prolonged power outage. Thus, it is more sensitive particularly for subtypes A and A/G HIV-1 in resource-limited settings.