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Journal of Clinical Microbiology, October 1998, p. 2964-2969, Vol. 36, No. 10
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Ultrasensitive Reverse Transcription-PCR Assay for Quantitation
of Human Immunodeficiency Virus Type 1 RNA in Plasma
Rita
Sun,1
Joanne
Ku,1
Harsha
Jayakar,1
Jo-Chi
Kuo,1
Donald
Brambilla,2
Steven
Herman,1
Maurice
Rosenstraus,1,* and
Joanne
Spadoro1
Roche Molecular Systems, Inc., Branchburg,
New Jersey 08876,1 and
New England
Research Institutes, Watertown, Massachusetts 021722
Received 4 March 1998/Returned for modification 15 May
1998/Accepted 30 June 1998
With the recent introduction of combination therapy, human
immunodeficiency virus type 1 (HIV-1) RNA levels in plasma have been
dramatically reduced, frequently to below the limit of quantitation (400 copies/ml of plasma) of the AMPLICOR HIV-1 MONITOR Test (Roche Diagnostic Systems). To achieve enhanced sensitivity of the
AMPLICOR HIV-1 MONITOR Test, a modified specimen preparation procedure that allows input of RNA from 10-fold more plasma per
amplification reaction was developed. This "ultrasensitive" method
allows the accurate quantitation of plasma HIV-1 RNA levels as low as
50 copies/ml. A precision study yielded average within-run and
between-run coefficients of variation (CV) of 24.8 and 9.6%,
respectively. A multicenter reproducibility study demonstrated that the
laboratory-to-laboratory reproducibility of this assay is good, with an
average CV of 32%. The linear range of this test is between 50 and
50,000 copies/ml of plasma. RNA concentrations measured by the
ultrasensitive and standard HIV-1 MONITOR tests exhibited good
agreement within the shared linear range of the two methods. The two
measurements were within a factor of 2 for 91% of the specimens
tested, with the concentration measured by the ultrasensitive method
being only slightly lower (median, 22% lower). Preliminary studies
suggest that this assay will prove to be useful for predicting
the stability of viral suppression in patients whose RNA
levels drop below 400 copies/ml in response to highly active
antiretroviral therapy.
*
Corresponding author. Mailing address: Roche Molecular
Systems, 1080 Route 202, Somerville, NJ 08876. Phone: (908) 253-7463. Fax: (908) 253-3318. E-mail:
maurice.rosenstraus{at}roche.com.
Journal of Clinical Microbiology, October 1998, p. 2964-2969, Vol. 36, No. 10
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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