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. 2022:2407:365-372.
doi: 10.1007/978-1-0716-1871-4_24.

TILDA: Tat/Rev Induced Limiting Dilution Assay

Cynthia Lungu  1 , Francesco A Procopio  2
Affiliations

TILDA: Tat/Rev Induced Limiting Dilution Assay

Cynthia Lungu et al. Methods Mol Biol. 2022.

Abstract

Recently the Tat/rev Induced Limiting Dilution Assay, or TILDA, has been proposed as a possible alternative method to quantify the HIV-1 reservoir. TILDA estimates the frequency of latently infected cells by probing, in a limiting dilution format, the presence or inducibility of tat and rev multiply spliced HIV-1 RNA. In doing so, TILDA reduces overestimation of reservoir size compared to HIV-1 DNA measurements because multiply spliced HIV-1 RNA is less likely to be transcribed from dysfunctional genomes with replication defects. TILDA is easy to perform, requires a very low input number of cells and has a fast turnaround time, making it ideal for use in clinical settings. Here we describe the execution of TILDA with particular emphasis on cell preparation and the limiting dilution scheme.

Keywords: HIV latency; HIV reservoir quantification; Inducible reservoir; TILDA.

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