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Journal of Clinical Microbiology, April 2001, p. 1334-1338, Vol. 39, No. 4
0095-1137/01/$04.00+0 DOI: 10.1128/JCM.39.4.1334-1338.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Comparison of Quantitative and Qualitative PCR
Assays for Cytomegalovirus DNA in Plasma
Angela M.
Caliendo,1,*
Rob
Schuurman,2
Belinda
Yen-Lieberman,3
Stephen A.
Spector,4
Janet
Andersen,5
R.
Manjiry,2
Clyde
Crumpacker,6
Nell S.
Lurain,7 and
Alejo
Erice8 for the Cmv Working
Group of the Complications of Hiv Disease
Rac, Aids Clinical Trials Group
Department of Pathology and Laboratory Medicine and
Department of Medicine, Division of Infectious Diseases, Emory
University, Atlanta, Georgia1;
Department of Virology, Eykman Winkler Institute for Clinical
Microbiology, Utrecht University Hospital, Utrecht, The
Netherlands2; Department of Clinical
Pathology, Cleveland Clinic Foundation, Cleveland,
Ohio3; Department of Pediatric
Infectious Diseases, University of California San Diego, San Diego,
California4; Department of
Immunology/Microbiology, Statistics and Data Analysis Center,
Harvard School of Public Health,5 and
Division of Infectious Diseases, Beth Israel Deaconess
Medical Center, Harvard Medical School,6 Boston,
Massachusetts; Department of Immunology/Microbiology,
Rush-Presbyterian-St. Luke's Medical Center, Chicago,
Illinois7; and Division of Infectious
Diseases, Department of Laboratory Medicine and Pathology and
Department of Medicine, University of Minnesota Medical School,
Minneapolis, Minnesota8
Received 11 August 2000/Returned for modification 13 December
2000/Accepted 22 January 2001
We analyzed the performance characteristics of the qualitative
AMPLICOR CMV Test (Roche Molecular Systems, Pleasanton, Calif.) and
quantitative COBAS AMPLICOR CMV MONITOR Test (Roche Molecular Systems)
assays and compared the performance of the AMPLICOR quantitative assay
with an in-house-developed cytomegalovirus (CMV) DNA PCR assay. The
quantitative AMPLICOR assay was found to be more sensitive than the
qualitative AMPLICOR assay. The quantitative AMPLICOR assay has a lower
limit of sensitivity of 400 CMV DNA copies/ml of plasma and is linear
to 50,000 CMV DNA copies/ml of plasma. Compared to the in-house PCR
assay, the AMPLICOR quantitative assay gave lower viral load values at
all concentrations tested, but the difference between the two assays
was not consistent across the entire dynamic range of the AMPLICOR
quantitative assay. At the lower end of the assay, the viral load
values obtained with the in-house PCR assay were three- to fivefold
(0.5 to 0.7 log units) higher than those measured with the AMPLICOR
assay. At higher input concentrations, the differences between the two
assays approached 10-fold. This direct comparison of the in-house assay and the quantitative AMPLICOR assay provides the ability to compare previously published in-house data with an assay widely available for
future research and clinical monitoring of patients with CMV infections.
*
Corresponding author. Mailing address: Emory University
Hospital, Clinical Laboratory, F147, 1364 Clifton Rd., NE, Atlanta, GA
30322. Phone: (404) 712-5721. Fax: (404) 712-4632. E-mail: acalien{at}emory.edu.
Journal of Clinical Microbiology, April 2001, p. 1334-1338, Vol. 39, No. 4
0095-1137/01/$04.00+0 DOI: 10.1128/JCM.39.4.1334-1338.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
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