PT  - JOURNAL ARTICLE
AU  - Christian Gaebler
AU  - Julio C. C. Lorenzi
AU  - Thiago Y. Oliveira
AU  - Lilian Nogueira
AU  - Victor Ramos
AU  - Ching-Lan Lu
AU  - Joy A. Pai
AU  - Pilar Mendoza
AU  - Mila Jankovic
AU  - Marina Caskey
AU  - Michel C. Nussenzweig
TI  - Quadruplex qPCR for qualitative and quantitative analysis of the HIV-1 latent reservoir
AID  - 10.1101/641951
DP  - 2019 Jan 01
TA  - bioRxiv
PG  - 641951
4099  - http://biorxiv.org/content/early/2019/05/31/641951.short
4100  - http://biorxiv.org/content/early/2019/05/31/641951.full
AB  - HIV-1 infection requires life-long therapy with anti-retroviral 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 immunologic 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 polymerase chain reaction (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 2 of the 4 probes (Q4PCR). This sensitive and specific approach allows for an unbiased characterization of the HIV-1 latent reservoir.Summary HIV-1 cure research seeks to decrease or eliminate the latent reservoir. The evaluation of such curative strategies requires accurate measures of the reservoir. Gaebler et al. describe a combined multicolor qPCR and next generation sequencing method that enables the sensitive and specific characterization of the HIV-1 latent reservoir.