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Journal of Clinical Microbiology, June 1999, p. 1813-1818, Vol. 37, No. 6
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Design and Evaluation of a Human Immunodeficiency Virus Type 1 RNA Assay Using Nucleic Acid Sequence-Based Amplification Technology Able To Quantify Both Group M and O Viruses by Using the Long Terminal Repeat as Target

Michel P. de Baar,1,* Audrey M. van der Schoot,1 Jaap Goudsmit,1 Femke Jacobs,2 Ron Ehren,2 Karin H. M. van der Horn,1 Peter Oudshoorn,2 Frank de Wolf,1 and Anthony de Ronde1

Department of Human Retrovirology, Academic Medical Center, University of Amsterdam, Amsterdam,1 and Organon Teknika B.V., Boxtel,2 The Netherlands

Received 14 December 1998/Returned for modification 28 January 1999/Accepted 1 March 1999

Currently available human immunodeficiency virus type 1 (HIV-1) RNA quantification assays can detect most viruses of the group M subtypes, but a substantial number are missed or not quantified reliably. Viruses of HIV-1 group O cannot be detected by any commercially available assay. We developed and evaluated a quantitative assay based on nucleic acid sequence-based amplification (NASBA) technology, with primers and probes located in the conserved long terminal repeat (LTR) region of the HIV-1 genome. In 68 of 72 serum samples from individuals infected with HIV-1 subtypes A to H of group M, viruses could be detected and quantified. In serum samples from two patients infected with HIV-1 group O viruses, these viruses as well could be detected and quantified. In contrast, the currently used gag-based assay underestimated the presence of subtype A viruses and could not detect subtype G and group O viruses. The discrepancy between the results of the two assays may be explained by the number of mismatches found within and among the probe and primer regions of the subtype isolates. These data indicate that LTR-based assays, including the NASBA format chosen here, are better suited to monitoring HIV-1 therapy than are gag-based assays in an era in which multiple HIV-1 subtypes and groups are spreading worldwide.

* Corresponding author. Mailing address: Department of Human Retrovirology, Academic Medical Center, University of Amsterdam, Meibergdreef 15, 1105 AZ Amsterdam, The Netherlands. Phone: 31-20-566 8569. Fax: 31-20-691 6531. E-mail: M.P.deBaar{at}amc.uva.nl.

Journal of Clinical Microbiology, June 1999, p. 1813-1818, Vol. 37, No. 6
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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