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Comparative Study
. 2005 Jan;43(1):293-8.
doi: 10.1128/JCM.43.1.293-298.2005.

Evaluation of the COBAS TaqMan HCV test with automated sample processing using the MagNA pure LC instrument

Jeffrey J Germer  1 , W Scott Harmsen, Jayawant N Mandrekar, P Shawn Mitchell, Joseph D C Yao
Affiliations
Comparative Study

Evaluation of the COBAS TaqMan HCV test with automated sample processing using the MagNA pure LC instrument

Jeffrey J Germer et al. J Clin Microbiol. 2005 Jan.

Abstract

The COBAS TaqMan HCV Test (TaqMan HCV; Roche Molecular Systems Inc., Branchburg, N.J.) for hepatitis C virus (HCV) performed on the COBAS TaqMan 48 Analyzer (Roche Molecular Systems) currently relies on a manual sample processing method. Implementation of an automated sample processing method would facilitate the clinical use of this test. In this study, we evaluated the performance characteristics of TaqMan HCV following automated sample processing by the MagNA Pure LC instrument (MP; Roche Applied Science, Indianapolis, Ind.). The analytical sensitivity of TaqMan HCV following sample processing by MP was 8.1 IU/ml (95% confidence interval, 6.1 to 15.2). The assay showed good linearity (R(2) = 0.99) across a wide range of HCV RNA levels (25 to 5 x 10(6) IU/ml), with coefficients of variation ranging from 10% to 46%. Among 83 clinical specimens, the sensitivity and specificity of TaqMan HCV were 100% and 95%, respectively, when compared to the COBAS AMPLICOR hepatitis C virus test, version 2.0 (COBAS AMPLICOR; Roche Molecular Systems), with TaqMan HCV detecting two more HCV RNA-positive specimens than COBAS AMPLICOR. Both specimens were confirmed to be HCV RNA positive by the VERSANT HCV RNA qualitative test (Bayer HealthCare LLC, Tarrytown, N.Y.). There was also strong correlation (R(2) = 0.95) and good agreement between the results from TaqMan HCV and the VERSANT HCV RNA 3.0 assay (bDNA) (Bayer HealthCare LLC) among a group of 93 clinical specimens. The MP is a versatile, labor-saving sample processing platform suitable for reliable performance of TaqMan HCV.

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Figures

FIG. 1.
FIG. 1.
Correlation between observed and expected results of HCV standards tested by TaqMan HCV combined with sample processing by MP.
FIG. 2.
FIG. 2.
Correlation between the results of 93 clinical specimens tested by TaqMan HCV combined with sample processing by MP and bDNA. HCV genotypes: (□) genotype 1; (▵) genotype 2; (×) genotype 3; (⋄) genotype 4; (▿) genotype 5; (star) genotype 6; and (boxed X) unable to genotype.
FIG. 3.
FIG. 3.
Log10 differences in quantitation between TaqMan HCV combined with sample processing by MP and bDNA among 93 clinical specimens (Bland-Altman plot). HCV genotypes: (□) genotype 1; (▵) genotype 2; (×) genotype 3; (⋄) genotype 4; (▿) genotype 5; (star) genotype 6; and (boxed X) unable to genotype.
FIG. 4.
FIG. 4.
Comparison of automated (MP) and manual (High Pure) sample processing workflow. The bars represent the general workflow for each sample processing method extending from sample preparation through completion of K-tube loading. The hands-on time, hands-off time, and total time required for the processing of 24 samples (21 specimens and 3 controls) by each method are also indicated (in minutes).
See this image and copyright information in PMC

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