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Journal of Clinical Microbiology, June 2007, p. 1723-1727, Vol. 45, No. 6
0095-1137/07/$08.00+0     doi:10.1128/JCM.02558-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Evaluation of Real-Time PCR Laboratory-Developed Tests Using Analyte-Specific Reagents for Cytomegalovirus Quantification{triangledown}

Angela M. Caliendo,1,2* Jessica Ingersoll,1 Andrea M. Fox-Canale,3 Sabine Pargman,1 Tameka Bythwood,1 Mary K. Hayden,4 James W. Bremer,3 and Nell S. Lurain3

Department of Pathology and Laboratory Medicine, Emory University School of Medicine,1 Center for AIDS Research, Emory University, Atlanta, Georgia,2 Department of Immunology/Microbiology,3 Section of Infectious Diseases, Rush University Medical Center, Chicago, Illinois4

Received 20 December 2006/ Returned for modification 5 March 2007/ Accepted 23 March 2007

Viral load testing for cytomegalovirus (CMV) has become the standard for the diagnosis of infection and monitoring of therapy at many transplant centers. However, no viral load test has been approved by the FDA. Therefore, many laboratories rely on laboratory-developed assays. This study evaluated the performance characteristics of two real-time PCR tests developed using the artus CMV analyte-specific reagents (ASRs). One version is distributed by Abbott Molecular and the other by QIAGEN. For plasma specimens, the Abbott test had a limit of detection of 2.3 log10 copies/ml and a linear range up to at least 6.0 log10 copies/ml. Comparison of plasma viral loads using the Abbott test and the Roche Amplicor Monitor test showed a mean difference of –0.012 log10 copies/ml. In addition, the Abbott test viral loads correlated with the Digene Hybrid Capture assay ratios. Viral loads obtained from plasma specimens tested by the Abbott and QIAGEN tests were in very close agreement (mean difference, 0.144 log10 copies/ml). When the QIAGEN test was evaluated with the QIAGEN, MagNA Pure, and easyMAG extraction methods, the viral loads for all three methods were within 0.370 log10 copies/ml. Thus, there is good agreement between viral loads obtained by the different tests using the same extraction method or by the same test using different extraction methods. The availability of real-time PCR ASRs provides additional reagents that can be used for CMV viral load testing.

* Corresponding author. Mailing address: Clinical Laboratories, H180, Emory University Hospital, 1364 Clifton Road, Atlanta, GA 30322. Phone: (404) 712-5721. Fax: (404) 727-3133. E-mail: acalien{at}emory.edu

{triangledown} Published ahead of print on 4 April 2007.

Journal of Clinical Microbiology, June 2007, p. 1723-1727, Vol. 45, No. 6
0095-1137/07/$08.00+0     doi:10.1128/JCM.02558-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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