Molecular confirmation of HIV-1 and HIV-2 coinfections among initially serologically dually-reactive samples from patients living in West Africa

PLOS ONE RESEARCH ARTICLE Molecular confirmation of HIV-1 and HIV-2 coinfections among initially serologically dually-reactive samples from patients living in West Africa a1111111111 a1111111111 a1111111111 a1111111111 a1111111111 OPEN ACCESS Boris K. Tchounga1,2, Mélanie Bertine3, Florence Damond3, Valentine Marie Ferré3, André Inwoley4,5, Simon P. Boni1, Alice Moisan6, Jean-Christophe Plantier6, Diane Descamps3, Didier K. Ekouevi ID1,7,8,9*, Charlotte Charpentier3 1 Programme PACCI, Site ANRS, Abidjan, Côte d’Ivoire, 2 Elizabeth Glaser Pediatric AIDS Foundation, Yaoundé, Cameroun, 3 Service de Virologie, Université Paris Cité, INSERM, IAME, UMR 1137, AP-HP, Hôpital Bichat-Claude Bernard, Paris, France, 4 Félix Houphouet-Boigny University, Abidjan, Côte d’Ivoire, 5 Research and Diagnosis Center for AIDS and Other Infectious Diseases (CeDReS), CHU (University Hospital) of Treichville, Abidjan, Côte d’Ivoire, 6 Normandie Univ., UNIROUEN, EA2656, GRAM, CHU de Rouen, Laboratoire de Virologie associé au CNR du VIH, Rouen, France, 7 ISPED, Université de Bordeaux & Centre INSERM U1219—Bordeaux Population Health, Bordeaux, France, 8 Université de Lomé, Faculté des Sciences de la Santé, Département de Santé Publique, Lomé, Togo, 9 Centre Africain de Recherche en Epidémiologie et en Santé Publique (CARESP), Lomé, Togo * Citation: Tchounga BK, Bertine M, Damond F, Ferré VM, Inwoley A, Boni SP, et al. (2023) Molecular confirmation of HIV-1 and HIV-2 coinfections among initially serologically duallyreactive samples from patients living in West Africa. PLoS ONE 18(3): e0283602. https://doi.org/ 10.1371/journal.pone.0283602 Abstract Editor: Jean-Pierre Vartanian, Institut Pasteur, FRANCE This study aimed to confirm the co-infection with HIV-1 and HIV-2, among West African patients using in-house HIV type/group enzyme-immuno assays and molecular diagnosis. Objectives Received: January 22, 2022 Accepted: March 13, 2023 Published: March 31, 2023 Copyright: © 2023 Tchounga et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Data Availability Statement: The data belong to the IDeA West-Africa consortium. The first author, corresponding author and others are members of IDEA West Africa, so they had the rights to access and use the data that other researchers outside the IDEA consortium have not (https://www.iedea.org/ regions/west-africa/; https://cutt.ly/o3UlYJU). Data can be made available to researchers outside the IDEA consortium upon request. The data can be accessed from Mr. Jean Claude Azany (). Design A cross-sectional survey was conducted from April 2016 to October 2017 in the biggest HIV clinics of Côte d’Ivoire and Burkina Faso. Method A first serological confirmation was done in the referral laboratory using an in-house, indirect immuno-enzymatic essay allowing the qualitative detection of both HIV-1 and HIV-2 antibodies. In order to separately detect anti-HIV-1 and anti-HIV-2 antibodies, a type/group specific enzyme-immuno assay (HIV-GSEIA) was used. To confirm the co-infections, HIV-1 and HIV-2 DNA-qualitative PCR assays were performed. Results A total of 91 patients were enrolled in the study and provided blood sample for HIV type confirmatory testing including 13 (14.3%) HIV-2 mono-reactive and 78 (85.7%) HIV-1/HIV-2 dually-reactive based on the HIV testing National Algorithms. The first serological ELISA confirmatory test performed showed that 80 (78.9%) of the 91 participants were dually- PLOS ONE | https://doi.org/10.1371/journal.pone.0283602 March 31, 2023 1/8 PLOS ONE Funding: This work was supported by The European & Developing Countries Clinical Trials Partnership (EDCTP), under the EDCTP2 Training and Mobility Action awarded to Dr BKT as Career Development Fellowship grant. [Grant number: TMA 2016 CDF 1597 EDIIMark-2; https://www. edctpgrants.org/publicportal#/search]. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. All funding sources have been reported for this study. There was no additional external funding received for this study. Molecular confirmation of HIV-1 and HIV-2 in West Africa reactive. The HIV-GSEIA performed on these 80 serum samples retrieve one 61 HIV-1/HIV2 dually-reactive samples. HIV-1 and HIV-2 DNA PCR were performed on 54 of the 61 HIV1/HIV-2 dually-reactive samples and 46 out of 61 (75.4%) samples were found HIV-1/HIV-2 coinfected. Conclusion The contribution of type/group specific enzyme-immuno assay to accurately identify HIV-1/ HIV-2 coinfections remain suboptimal, emphasizing the need for molecular diagnosis platforms in West Africa, to avail HIV DNA PCR test for the confirmation of HIV-1/HIV-2 coinfections. Competing interests: The authors have declared that no competing interests exist. Introduction West Africa is characterized by the co-circulation of HIV-1 and HIV-2, leading to co-infections with both viruses [1]. ART-naïve patients co-infected with both viruses were reported to experience a higher mortality rates compared to HIV-2 mono-infected patients [2, 3]. Moreover, providing long-term adequate antiretroviral (ARV) treatment to patients infected with both viruses is challenging in sub-Saharan African countries, where access to ARV drugs effective on patients living with HIV-2 and experiencing viral resistance to protease inhibitors (PI) and integrase inhibitors is still scarce [1, 4, 5]. The accurate diagnosis of this co-infection remains challenging for the national HIV testing algorithms of West African countries [6–8]. Currently, only molecular diagnosis methods consisting in the detection of HIV-1 and HIV-2 DNA in the same patients constitute a gold standard for the confirmation of HIV-1 and HIV-2 co-infections [6, 8]. Only one study described a cohort of 17 patients co-infected, with positive HIV-1 RNA and HIV-2 proviral DNA [9]. For most of the studies with no molecular diagnosis, there is persistent doubts on the accuracy of this diagnosis and the most appropriate expression to describe these patients is dually-reactive for HIV-1 and HIV-2. The main challenge faced by the West African countries is access to molecular technics for the accurate diagnosis of HIV-1 and HIV-2 co-infections. Some in-house molecular tests have been developed, validated and patented for the molecular diagnosis of both HIV-1 and HIV-2. However these in-house or manufactured tests are scarcely available for referral laboratories of West African countries [8]. The alternative remains the systematic confirmation of HIV-1 and HIV-2 dually-reactive results from rapid tests used in the national algorithm, with a more specific and sensitive in-house ELISA technic that can be affordable and routinely used in referral laboratories. The aim of (...truncated)