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Updated: Feb 15, 2026

Amplifying and Quantifying HIV-1 RNA in HIV Infected Individuals with Viral Loads Below the Limit of Detection by Standard Clinical Assays

Amplifying and Quantifying HIV-1 RNA in HIV Infected Individuals with Viral Loads Below the Limit of Detection by Standard Clinical Assays

Published on: September 26, 2011

Single-molecule techniques to quantify and genetically characterise persistent HIV.

Xiao Qian Wang¹, Sarah Palmer²

¹Centre for Virus Research, The Westmead Institute for Medical Research, The University of Sydney, 176 Hawkesbury Road, Westmead, NSW, 2145, Australia.

|January 11, 2018

Summary

This summary is machine-generated.

Single-molecule assays reveal persistent, low-level HIV-1 RNA in plasma and cellular reservoirs despite effective antiretroviral therapy. These methods enhance understanding of the HIV-1 reservoir dynamics and inform future eradication strategies.

Keywords:

Full-length individual proviral sequencing Intracellular HIV-1 reservoirs Persistent HIV-1 Single-copy assay Single-genome/proviral sequencing Single-molecule assays

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Rapid Screening of HIV Reverse Transcriptase and Integrase Inhibitors

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Published on: April 9, 2014

Area of Science:

Virology
Immunology
Molecular Biology

Background:

Antiretroviral therapy (ART) suppresses HIV-1 but does not eradicate it.
Persistent low-level HIV-1 RNA and proviruses remain detectable in plasma and cellular reservoirs during effective ART.
Cessation of ART leads to viral rebound, highlighting the need to understand persistent HIV-1 reservoirs.

Purpose of the Study:

To comprehensively examine the quantity and genetic composition of persistent HIV-1 in plasma and cellular reservoirs.
To elucidate the source and viral dynamics of persistent HIV-1 during long-term effective therapy.
To evaluate the utility of single-molecule techniques in understanding HIV-1 persistence.

Main Methods:

Single-molecule techniques, including single-copy assay (SCA) and single-genome/proviral sequencing (SGS).
SCA quantifies plasma HIV-1 RNA down to a single copy.
SGS genetically characterizes HIV-1 populations under various selective pressures.

Main Results:

Low-level HIV-1 RNA persists in plasma and cerebrospinal fluid (CSF) despite ART, treatment intensification, and other therapies.
The intracellular HIV-1 reservoir is stable, primarily residing in CD4+ memory T cells.
Cellular proliferation maintains the reservoir, with potential cryptic replication in sub-optimal treatment sites.
Latency-reversing agents activate quiescent proviruses but do not clear the reservoir.
Full-length sequencing reveals most intracellular HIV-1 DNA is defective; intact proviruses are unequally distributed.

Conclusions:

Single-molecule assays significantly enhance understanding of persistent HIV-1 dynamics in plasma and cells.
These techniques are crucial for future research into the HIV-1 reservoir and developing eradication strategies.
The intracellular reservoir is stable and maintained by cell proliferation, with genetic defects prevalent in proviral DNA.