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Immunology and Infection

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JoVE Journal Immunology and Infection
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

Article DOI: 10.3791/2960-v • 13:59 min • September 26th, 2011
September 26th, 2011 •
1, 1, 1, 1, 1, 2, 3
1The virology Core at the HIV Drug Resistance Program, NCI-Frederick, 2Division of Infectious Diseases, University of Pittsburgh, 3Department of Molecular Biology and Microbiology, Tuffts University

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Quantifying levels of HIV-1 RNA in plasma and sequencing single HIV-1 genomes from individuals with viral loads below the limit of detection (50-75 copies/ml) is difficult. Here we describe how to extract and quantify plasma viral RNA using a real time PCR assay that reliably measures HIV-1 RNA down to 0.3 copies/ml and how to amplify viral genomes by single genome sequencing, from samples with very low viral loads.

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Amplifying Quantifying HIV-1 RNA Viral Loads Limit Of Detection Standard Clinical Assays Viral Genes Insight Viral Dynamics Virus Host Interactions Patients Natural Control Combination Antiretroviral Therapy (cART) Single Genome Sequencing (SGS Protocol) Single-copy Assay (SCA Protocol) Real-time PCR Assay Sensitivity Volume Of Plasma RNA Copy Number Internal Control Testing DNA Contamination Triplicate Measurements Single-genome Sequencing Assay (SGS) Non-labor Intensive Limiting Dilution Assay Endpoint Diluted CDNA Product
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