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Wennett Post-Doctoral Fellowship"}],"content-domain":{"domain":["rupress.org"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2019,10,7]]},"abstract":"<jats:p>HIV-1 infection requires lifelong therapy with antiretroviral drugs due to the existence of a latent reservoir of transcriptionally inactive integrated proviruses. The goal of HIV-1 cure research is to eliminate or functionally silence this reservoir. To this end, there are numerous ongoing studies to evaluate immunological approaches, including monoclonal antibody therapies. Evaluating the results of these studies requires sensitive and specific measures of the reservoir. Here, we describe a relatively high-throughput combined quantitative PCR (qPCR) and next-generation sequencing method. Four different qPCR probes covering the packaging signal (PS), group-specific antigen (gag), polymerase (pol), and envelope (env) are combined in a single multiplex reaction to detect the HIV-1 genome in limiting dilution samples followed by sequence verification of individual reactions that are positive for combinations of any two of the four probes (Q4PCR). This sensitive and specific approach allows for an unbiased characterization of the HIV-1 latent reservoir.<\/jats:p>","DOI":"10.1084\/jem.20190896","type":"journal-article","created":{"date-parts":[[2019,7,26]],"date-time":"2019-07-26T15:05:34Z","timestamp":1564153534000},"page":"2253-2264","update-policy":"https:\/\/doi.org\/10.1084\/jem.crossmarkpolicy","source":"Crossref","is-referenced-by-count":125,"title":["Combination of quadruplex qPCR and next-generation sequencing for qualitative and quantitative analysis of the HIV-1 latent reservoir"],"prefix":"10.1084","volume":"216","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7295-8128","authenticated-orcid":true,"given":"Christian","family":"Gaebler","sequence":"first","affiliation":[{"name":"Laboratory of Molecular Immunology, The Rockefeller University, New York, NY 1"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julio C.C.","family":"Lorenzi","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Immunology, The Rockefeller University, New York, NY 1"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thiago Y.","family":"Oliveira","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Immunology, The Rockefeller University, New York, NY 1"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8168-2629","authenticated-orcid":true,"given":"Lilian","family":"Nogueira","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Immunology, The Rockefeller University, New York, NY 1"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7353-3420","authenticated-orcid":true,"given":"Victor","family":"Ramos","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Immunology, The Rockefeller University, New York, NY 1"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ching-Lan","family":"Lu","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Immunology, The Rockefeller University, New York, NY 1"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3580-8061","authenticated-orcid":true,"given":"Joy A.","family":"Pai","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Immunology, The Rockefeller University, New York, NY 1"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pilar","family":"Mendoza","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Immunology, The Rockefeller University, New York, NY 1"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9696-5234","authenticated-orcid":true,"given":"Mila","family":"Jankovic","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Immunology, The Rockefeller University, New York, NY 1"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marina","family":"Caskey","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Immunology, The Rockefeller University, New York, NY 1"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0592-8564","authenticated-orcid":true,"given":"Michel C.","family":"Nussenzweig","sequence":"additional","affiliation":[{"name":"Laboratory of Molecular Immunology, The Rockefeller University, New York, NY 1"},{"name":"Howard Hughes Medical Institute, The Rockefeller University, New York, NY 2"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"291","published-online":{"date-parts":[[2019,7,26]]},"reference":[{"key":"2023072609365473700_bib1","doi-asserted-by":"publisher","first-page":"151","DOI":"10.1016\/j.jviromet.2010.01.012","article-title":"Rational design of HIV-1 fluorescent hydrolysis probes considering phylogenetic variation and probe performance","volume":"165","author":"Althaus","year":"2010","journal-title":"J. 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