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Journal of Clinical Microbiology, August 1999, p. 2557-2563, Vol. 37, No. 8
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Development of Calibrated Viral Load Standards for Group M Subtypes of Human Immunodeficiency Virus Type 1 and Performance of an Improved AMPLICOR HIV-1 MONITOR Test with Isolates of Diverse Subtypes

Nelson L. Michael,1,* Steven A. Herman,2 Shirley Kwok,3 Kimberly Dreyer,2,dagger June Wang,2 Cindy Christopherson,3 Joanne P. Spadoro,2 Karen K. Y. Young,2 Victoria Polonis,4 Francine E. McCutchan,4 Jean Carr,4 John R. Mascola,1,5 Linda L. Jagodzinski,4 and Merlin L. Robb1

Division of Retrovirology, Walter Reed Army Institute of Research,1 and Henry M. Jackson Foundation,4 Rockville, Maryland 20850; Diagnostics Development, Roche Molecular Systems, Somerville, New Jersey 088762; Discovery Research, Roche Molecular Systems, Alameda, California 945013; and Department of Infectious Diseases, Naval Medical Research Institute, Bethesda, Maryland 208895

Received 28 December 1998/Returned for modification 15 March 1999/Accepted 10 May 1999

Accurate determination of plasma human immunodeficiency virus type 1 (HIV-1) RNA levels is critical for the effective management of HIV-1 disease. The AMPLICOR HIV-1 MONITOR Test, a reverse transcription-PCR-based test for quantification of HIV-1 RNA in plasma, was developed when little sequence information on HIV-1 isolates from outside North America was available. It has since become apparent that many non-subtype B isolates, particularly subtypes A and E, are detected inefficiently by the test. We describe here the AMPLICOR HIV-1 MONITOR Test, version 1.5, an upgraded test developed to minimize subtype-related variation. We also developed a panel of HIV-1 standards containing 30 HIV-1 isolates of subtypes A through G. The virus particle concentration of each cultured viral stock was standardized by electron microscopic virus particle counting. We used this panel to determine the performance of the original AMPLICOR HIV-1 MONITOR Test and version 1.5 of the test with HIV-1 subtypes A through G. The original test underestimated the concentration of HIV-1 subtype A, E, F, and G RNA by 10-fold or more, whereas version of the 1.5 test yielded equivalent quantification of HIV-1 RNA regardless of the subtype. In light of the increasing intermixing of HIV-1 subtypes worldwide, standardization of PCR-based tests against well-characterized viral isolates representing the full range of HIV-1 diversity will be essential for the continued utility of these important clinical management tools.

* Corresponding author. Mailing address: Department of Molecular Diagnostics and Pathogenesis, Division of Retrovirology, Walter Reed Army Institute of Research, 1600 E. Gude Dr., Rockville, MD 20850. Phone: (301) 762-0089, ext. 1081. Fax: (301) 762-7460. E-mail: nmichael{at}pasteur.hjf.org.

dagger Present address: Department of Infectious Diseases, University of Rochester Medical Center, Rochester, NY 14627.

Journal of Clinical Microbiology, August 1999, p. 2557-2563, Vol. 37, No. 8
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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