Specific quantification of inducible HIV-1 reservoir by RT-LAMP

通过 RT-LAMP 对可诱导 HIV-1 病毒库进行特异性定量

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作者:Tanvir Hossain #, Cynthia Lungu #, Sten de Schrijver, Mamokoena Kuali, Raquel Crespo, Nicole Reddy, Ayanda Ngubane, Tsung Wai Kan, Kavidha Reddy, Shringar Rao, Robert-Jan Palstra, Paradise Madlala, Thumbi Ndung'u, Tokameh Mahmoudi

Background

Strategies toward HIV-1 cure

Conclusions

SQuHIVLa offers a high throughput, scalable, and specific HIV-1 reservoir quantification tool that is amenable to resource-limited settings. This assay poses remarkable potential in facilitating the evaluation of potential interventional strategies toward achieving HIV-1 cure.

Methods

We developed Specific Quantification of Inducible HIV-1 by RT-LAMP (SQuHIVLa), leveraging the high sensitivity and specificity of RT-LAMP, performed in a single reaction, to detect and quantify cells expressing tat/rev HIV-1 multiply spliced RNA (msRNA) upon activation. The LAMP primer/probe used in SQuHIVLa was designed to exclusively detect HIV-1 tat/rev msRNA and adapted for different HIV-1 subtypes.

Results

Using SQuHIVLa, we successfully quantify the inducible viral reservoir in CD4+ T cells from people living with HIV-1 subtypes B and C on cART. The assay demonstrates high sensitivity, specificity, and reproducibility. Conclusions: SQuHIVLa offers a high throughput, scalable, and specific HIV-1 reservoir quantification tool that is amenable to resource-limited settings. This assay poses remarkable potential in facilitating the evaluation of potential interventional strategies toward achieving HIV-1 cure.

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