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Journal of Clinical Microbiology, August 2003, p. 3503-3508, Vol. 41, No. 8
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.8.3503-3508.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Evaluation of the MagNA Pure LC Instrument for Extraction of Hepatitis C Virus RNA for the COBAS AMPLICOR Hepatitis C Virus Test, Version 2.0

Jeffrey J. Germer,1 Michele M. Lins,1 Monica E. Jensen,1 W. Scott Harmsen,2 Duane M. Ilstrup,2 P. Shawn Mitchell,1 Franklin R. Cockerill III,1 and Robin Patel1,3*

Division of Clinical Microbiology,1 Section of Biostatistics,2 Division of Infectious Diseases, Mayo Clinic, Rochester, Minnesota 559063

Received 23 December 2002/ Returned for modification 13 March 2003/ Accepted 10 May 2003

The COBAS AMPLICOR system has played a major role in the transition of molecular diagnostics from research to routine clinical laboratory use by automating the nucleic acid amplification and detection processes. However, sample preparation remains a labor-intensive portion of the procedure. In this study, we evaluated the performance of the COBAS AMPLICOR Hepatitis C Virus Test, version 2.0 (Roche Molecular Systems, Branchburg, N.J.) following manual hepatitis C virus (HCV) RNA extraction versus automated extraction with the MagNA Pure LC instrument (Roche Applied Science, Indianapolis, Ind.). Parallel replicate testing was performed with standard dilutions of 100, 75, 60, and 0 HCV IU/ml and 153 clinical specimens. An analytical sensitivity of 75 IU/ml was achieved with either the manual or the standard-volume (200 µl) automated extraction methodologies (25 of 26 [96.2%]; 95% confidence interval [95% CI], 80.4 to 99.9), whereas the clinical sensitivity and specificity were both 100% with either extraction method. A large-volume (1 ml) automated extraction method was also evaluated with standard dilutions of 40, 25, 10, and 0 IU/ml and the same 153 clinical specimens. The analytical sensitivity of the COBAS AMPLICOR assay with the large-volume extraction method was 25 HCV IU/ml (26 of 26 [100%]; 95% CI, 86.8 to 100), whereas the clinical sensitivity and specificity were both 100%. The MagNA Pure LC instrument is a versatile, labor-saving platform capable of integration with minimal modification of the existing assay procedure. The increased sensitivity of the COBAS AMPLICOR Hepatitis C Virus Test, version 2.0 performed in conjunction with large-volume HCV RNA extraction may be important in HCV diagnostic testing as new therapeutic strategies evolve.

* Corresponding author. Mailing address: Division of Infectious Diseases, Mayo Clinic, 200 First St., SW, Rochester, MN 55905. Phone: (507) 255-6482. Fax: (507) 255-7767. E-mail: patel.robin{at}mayo.edu.

Journal of Clinical Microbiology, August 2003, p. 3503-3508, Vol. 41, No. 8
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.8.3503-3508.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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