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Journal of Clinical Microbiology, May 1999, p. 1260-1264, Vol. 37, No. 5
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Highly Sensitive Methods for Quantitation of Human Immunodeficiency Virus Type 1 RNA from Plasma, Cells, and Tissues

Marek Fischer,1,* Werner Huber,1,dagger Alex Kallivroussis,1 Peter Ott,2 Milos Opravil,1 Ruedi Lüthy,1,Dagger Rainer Weber,1 and Richard W. Cone1

Division of Infectious Diseases and Hospital Epidemiology, Department of Medicine,1 and Clinic for Ear, Nose and Throat Surgery,2 University Hospital Zurich, CH-8091 Zurich, Switzerland

Received 28 August 1998/Returned for modification 9 December 1998/Accepted 26 January 1999

Precise and sensitive quantitation of viral RNA in specimens from human immunodeficiency virus (HIV) type 1 (HIV-1)-infected individuals has become an indispensable tool for the monitoring of the efficacy of highly active antiretroviral combination therapy. The present report describes reproducible and efficient protocols with enhanced sensitivity for quantitation of HIV-1 RNA from plasma, peripheral blood mononuclear cells, and tissues with Qiagen silica columns for RNA purification combined with the Roche Amplicor HIV-1 Monitor test for quantitative reverse transcription-PCR (RT-PCR). Extraction of RNA from 0.5 ml of plasma resulted in the detection of fewer than 20 HIV RNA copies/ml of plasma, equivalent to the centrifugation-based boosted RT-PCR assay. Silica extraction of cellular RNA resulted in the detection of fewer than 3 HIV-1 RNA copies/µg of total RNA. These techniques facilitate direct comparisons of viral loads between liquid and cellular specimens. Application of these sensitive methods may improve the assessment of the response to new antiretroviral regimens.

* Corresponding author. Mailing address: University Hospital, Division of Infectious Diseases, Raemistrasse 100, CH-8091 Zurich, Switzerland. Phone: 41-1-255-3610. Fax: 41-1-255-3291. E-mail: inffia{at}usz.unizh.ch.

dagger Present address: Swiss Serum and Vaccine Institute Bern, CH-3001 Bern, Switzerland.

Dagger Present address: Carmenstrasse 23, CH-8032 Zurich, Switzerland.

Journal of Clinical Microbiology, May 1999, p. 1260-1264, Vol. 37, No. 5
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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