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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,
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.
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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