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Journal of Clinical Microbiology, July 2000, p. 2688-2695, Vol. 38, No. 7
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Evaluation of Performance of the Gen-Probe Human Immunodeficiency Virus Type 1 Viral Load Assay Using Primary Subtype A, C, and D Isolates from Kenya

Sandra Emery,1,2 Sharon Bodrug,3 Barbra A. Richardson,4 Cristina Giachetti,3 Martha A. Bott,3 Dana Panteleeff,1 Linda L. Jagodzinski,5 Nelson L. Michael,6 Ruth Nduati,7 Job Bwayo,8 Joan K. Kreiss,2,9 and Julie Overbaugh1,*

Division of Human Biology, Fred Hutchinson Cancer Research Center,1 and Departments of Biostatistics,4 Medicine,2 and Epidemiology,9 University of Washington, Seattle, Washington; Gen-Probe Incorporated, San Diego, California3; Henry M. Jackson Foundation,5 and Division of Retrovirology, Walter Reed Army Institute of Research,6 Rockville, Maryland; and Departments of Pediatrics7 and Medical Microbiology,8 University of Nairobi, Nairobi, Kenya

Received 11 November 1999/Returned for modification 13 March 2000/Accepted 24 April 2000

Accurate and sensitive quantification of human immunodeficiency virus type 1 (HIV-1) RNA has been invaluable as a marker for disease prognosis and for clinical monitoring of HIV-1 disease. The first generation of commercially available HIV-1 RNA tests were optimized to detect the predominant HIV-1 subtype found in North America and Europe, subtype B. However, these tests are frequently suboptimal in detecting HIV-1 genetic forms or subtypes found in other parts of the world. The goal of the present study was to evaluate the performance of a new viral load assay with non-subtype B viruses. A transcription-mediated amplification method for detection and quantitation of diverse HIV-1 subtypes, called the Gen-Probe HIV-1 viral load assay, is under development. In this study we examined the performance of the Gen-Probe HIV-1 viral load assay relative to that of the commonly used commercial HIV-1 RNA assays using a panel of primary isolates from Kenya. For comparison, we included several subtype B cloned viruses, and we quantified each virus using an in-house quantitative-competitive reverse transcriptase PCR (QC-RT-PCR) method and gagp24 antigen capture. The Gen-Probe HIV-1 viral load assay and a version of the Roche AMPLICOR HIV-1 MONITOR test (version 1.5) that was designed to detect a broader range of subtypes were both sensitive for the quantification of Kenyan primary isolates, which represented subtype A, C, and D viruses. The Gen-Probe HIV-1 viral load assay was more sensitive for the majority of viruses than the Roche AMPLICOR HIV-1 MONITOR test version 1.0, the Bayer Quantiplex HIV RNA 3.0 assay, or a QC-RT-PCR method in use in our laboratory, suggesting that it provides a useful method for quantifying HIV-1 RNAs from diverse parts of the world, including Africa.

* Corresponding author. Mailing address: Fred Hutchinson Cancer Research Center, 1100 Fairview Ave. N., C3-168, Seattle, WA 98109. Phone: (206) 667-3524. Fax: (206) 667-1535. E-mail: joverbau{at}fhcrc.org.

Journal of Clinical Microbiology, July 2000, p. 2688-2695, Vol. 38, No. 7
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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