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PMID: 15262500 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Quantification of infectious HIV-1 plasma viral load using a boosted in vitro infection protocol.

Virology ·Vol. 326 ·No. 1 ·2004-08-15 ·Pages 113-29

Rusert P, Fischer M, Joos B, Leemann C, Kuster H, Flepp M, Bonhoeffer S, Günthard HF, Trkola A

Abstract

Methods currently used for HIV-1 viral load measurements are very sensitive, but cannot distinguish between infectious and noninfectious particles. Here we describe the development of a novel, sensitive, and highly reproducible method that allows rapid isolation and quantification of infectious particles from patient plasma. By immobilizing HIV-1 particles in human plasma to platelets using polybrene, we observed a 10- to 1000-fold increase in infectivity over infection protocols using free virus particles. Using this method, we evaluated infectivity in plasma from 52 patients at various disease stages. At plasma viral loads of 1000-10000 HIV-1 RNA copies/ml 18%, at 10,000-50,000 copies/ml 73%, at 50,000-100,000 copies/ml 90%, and above 100,000 copies 96% of cultures were positive. We found that infectious titers among patients vary distinctively but are characteristic for a patient over extended time periods. Furthermore, we demonstrate that by evaluating infectious titers in conjunction with total HIV RNA loads, subtle effects of treatment intervention on viremia levels can be detected. The immobilization procedure does not interfere with viral entry and does not restore the infectivity of neutralized virus. Therefore, this assay system can be utilized to investigate the influence of substances that specifically affect virion infectivity such as neutralizing antibodies, soluble CD4, or protease inhibitors. Measuring viral infectivity may thereby function as an additional, useful marker in monitoring disease progression and evaluating efficacy of antivirals in vivo.

MeSH Terms

Adult Aged Anti-Retroviral Agents/therapeutic use Blood Platelets/virology Cells, Cultured Female HIV Infections/blood,drug therapy,virology HIV-1/genetics,isolation & purification,pathogenicity Hexadimethrine Bromide Humans Male Middle Aged Molecular Sequence Data Plasma/virology Polymerase Chain Reaction RNA, Viral/analysis Sensitivity and Specificity Viral Load/methods Virion/genetics

Chemicals

Anti-Retroviral Agents RNA, Viral Hexadimethrine Bromide

Authors & Affiliations

9 authors, click to expand affiliations / ORCID

Rusert Peter

Division of Infectious Diseases, University Hospital Zurich, Switzerland.

Fischer Marek

Joos Beda

Leemann Christine

Kuster Herbert

Flepp Markus

Bonhoeffer Sebastian

Günthard Huldrych F

Trkola Alexandra

Article Info

Journal

Virology

Abbr.

Virology

ISSN

0042-6822

DOI

10.1016/j.virol.2004.05.022

Published

2004-08-15

Pages

113-29

Language

English

Region

United States

NLM ID

0110674

Subset

Databases

GENBANK

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PubMed source DOI: 10.1016/j.virol.2004.05.022 View journal details

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Quantification of infectious HIV-1 plasma viral load using a boosted in vitro infection protocol.

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