{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,11]],"date-time":"2026-04-11T16:46:07Z","timestamp":1775925967196,"version":"3.50.1"},"reference-count":21,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2025,12,24]],"date-time":"2025-12-24T00:00:00Z","timestamp":1766534400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"},{"start":{"date-parts":[[2025,12,24]],"date-time":"2025-12-24T00:00:00Z","timestamp":1766534400000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/100005564","name":"Gilead Sciences","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100005564","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Journal of Medical Virology"],"published-print":{"date-parts":[[2026,1]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>Lenacapavir (LEN) is a first\u2010in\u2010class capsid inhibitor (CAI) that targets multiple stages of the HIV\u20101 lifecycle, showing efficacy in heavily treatment\u2010experienced (HTE) individuals with multidrug\u2010resistance (MDR) and in pre\u2010exposure prophylaxis (PrEP). This study aimed to characterize a novel in\u2010house next\u2010generation sequencing (NGS) protocol targeting HIV\u20101 capsid (CA) region using both HIV\u20101 RNA from plasma and HIV\u20101 DNA from peripheral\u2010blood\u2010mononuclear\u2010cells (PBMCs). A total of 60 samples (41 HIV\u20101 RNA and 19 HIV\u20101 DNA) with various HIV\u20101 subtypes and viremia levels were tested. Overall, molecular amplification was successful in 83.3% of cases (75.6% for HIV\u20101 RNA and 100% for HIV\u20101 DNA), while high quality sequences were obtained in 76.7% of samples (65.9% for HIV\u20101 RNA and 100% for HIV\u20101 DNA). Among RNA samples with viremia \u2265\u2009500 copies\/mL, sequencing success reached 92.6%, showing a statistically significant association with viral load. Subtype\u2010specific analysis showed amplification and sequencing rates of 86.0% and 79.1% for subtype B, and 76.5% and 70.6% for non\u2010B subtypes, with no significant difference. Reproducibility was fully confirmed by pairwise similarity analyses at 10% and 20% frequency cutoff, upon reprocessing 13 HIV\u20101 RNA samples. This protocol provides an important tool, primarily for subtype B, for personalized HIV\u20101 treatment with CAI\u2010based strategies, enabling efficient characterization of LEN resistance mutations in the CA region, using both DNA and RNA samples.<\/jats:p>","DOI":"10.1002\/jmv.70773","type":"journal-article","created":{"date-parts":[[2025,12,24]],"date-time":"2025-12-24T13:14:00Z","timestamp":1766582040000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Development of a Next\u2010Generation Sequencing Protocol for Assessing Lenacapavir Resistance in HIV\u20101 Capsid"],"prefix":"10.1002","volume":"98","author":[{"given":"Omar","family":"El Khalili","sequence":"first","affiliation":[{"name":"Department of Experimental Medicine University of Rome Tor Vergata Rome Italy"},{"name":"Ph.D. Course in Microbiology, Immunology, Infectious Diseases, and Transplants (MIMIT), University of Rome Tor Vergata Rome Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Collins Ambe","family":"Chenwi","sequence":"additional","affiliation":[{"name":"Department of Experimental Medicine University of Rome Tor Vergata Rome Italy"},{"name":"Ph.D. Course in Microbiology, Immunology, Infectious Diseases, and Transplants (MIMIT), University of Rome Tor Vergata Rome Italy"},{"name":"Chantal Biya International Reference Centre for Research on HIV and AIDS Prevention and Management (CIRCB)"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-4932-5344","authenticated-orcid":false,"given":"Daniele","family":"Spalletta","sequence":"additional","affiliation":[{"name":"Department of Experimental Medicine University of Rome Tor Vergata Rome Italy"},{"name":"Ph.D. Course in Microbiology, Immunology, Infectious Diseases, and Transplants (MIMIT), University of Rome Tor Vergata Rome Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Greta","family":"Marchegiani","sequence":"additional","affiliation":[{"name":"Department of Experimental Medicine University of Rome Tor Vergata Rome Italy"},{"name":"Ph.D. Course in Microbiology, Immunology, Infectious Diseases, and Transplants (MIMIT), University of Rome Tor Vergata Rome Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1713-8682","authenticated-orcid":false,"given":"Luca","family":"Carioti","sequence":"additional","affiliation":[{"name":"Department of Experimental Medicine University of Rome Tor Vergata Rome Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hossein Eizadi","family":"Moghadam","sequence":"additional","affiliation":[{"name":"Department of Experimental Medicine University of Rome Tor Vergata Rome Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ada","family":"Bertoli","sequence":"additional","affiliation":[{"name":"Department of Experimental Medicine University of Rome Tor Vergata Rome Italy"},{"name":"Virology Unit, Tor Vergata University Hospital Rome Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vincenzo","family":"Spagnuolo","sequence":"additional","affiliation":[{"name":"Infectious Diseases Unit, San Raffaele Scientific Institute Milan Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maria Mercedes","family":"Santoro","sequence":"additional","affiliation":[{"name":"Department of Experimental Medicine University of Rome Tor Vergata Rome Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Francesca","family":"Ceccherini\u2010Silberstein","sequence":"additional","affiliation":[{"name":"Department of Experimental Medicine University of Rome Tor Vergata Rome Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7717-2343","authenticated-orcid":false,"given":"Maria Concetta","family":"Bellocchi","sequence":"additional","affiliation":[{"name":"Department of Experimental Medicine University of Rome Tor Vergata Rome Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2025,12,24]]},"reference":[{"key":"e_1_2_12_2_1","unstructured":"Joint United Nations Programme on HIV\/AIDS (UNAIDS).2024. The Urgency of Now: AIDS at a Crossroads. Geneva: Joint United Nations Programme on HIV\/AIDS; 2024. 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WHO HIVResNet HIV Drug Resistance Laboratory Operational Framework. Second Edi. (WorldHealthOrganization ed.). Licence: CC BY\u2010NC\u2010SA 3.0 IGO.;2020."},{"key":"e_1_2_12_21_1","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/dkac388"},{"key":"e_1_2_12_22_1","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/dkae189"}],"container-title":["Journal of Medical Virology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/jmv.70773","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/jmv.70773","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/jmv.70773","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,4,11]],"date-time":"2026-04-11T16:13:23Z","timestamp":1775924003000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/jmv.70773"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,12,24]]},"references-count":21,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2026,1]]}},"alternative-id":["10.1002\/jmv.70773"],"URL":"https:\/\/doi.org\/10.1002\/jmv.70773","archive":["Portico"],"relation":{},"ISSN":["0146-6615","1096-9071"],"issn-type":[{"value":"0146-6615","type":"print"},{"value":"1096-9071","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,12,24]]},"assertion":[{"value":"2025-06-12","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-12-08","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-12-24","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e70773"}}