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Journal of Clinical Microbiology, February 2006, p. 318-323, Vol. 44, No. 2
0095-1137/06/$08.00+0     doi:10.1128/JCM.44.2.318-323.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Evaluation of the Invader Assay for Genotyping Hepatitis C Virus

Jeffrey J. Germer,1 David W. Majewski,1 Billy Yung,2 P. Shawn Mitchell,1 and Joseph D. C. Yao1*

Division of Clinical Microbiology, Mayo Clinic, Rochester, Minnesota,1 State University of New York Downstate Medical Center, Brooklyn, New York2

Received 10 January 2005/ Returned for modification 26 March 2005/ Accepted 23 November 2005

The Invader 1.0 assay (Invader HCV Genotyping Assay, version 1.0; Third Wave Technologies, Inc., Madison, WI) has been developed for the rapid differentiation of hepatitis C virus (HCV) genotypes 1 to 6 based on sequence variation within the HCV 5' noncoding (NC) region. In the present study, we evaluated the compatibility of Invader 1.0 with the COBAS MONITOR (COBAS AMPLICOR HCV MONITOR Test, version 2.0; Roche Molecular Systems, Inc., Branchburg, NJ), COBAS AMPLICOR (COBAS AMPLICOR Hepatitis C Virus Test, version 2.0; Roche Molecular Systems, Inc.), and COBAS TaqMan (COBAS TaqMan HCV Test; Roche Molecular Systems, Inc.) assays. The minimum HCV RNA titers required for successful HCV genotyping (≥90% success rate) were 1,000 IU/ml for COBAS MONITOR, 100 IU/ml for COBAS AMPLICOR, and 10 IU/ml for COBAS TaqMan. Invader 1.0 results obtained from unpurified COBAS TaqMan amplification products of 111 retrospectively selected clinical serum specimens (genotypes 1 to 6, with virus titers ranging from 15.1 to 2.1 x 107 IU/ml) showed 98% concordance with results obtained from the TRUGENE HCV 5' NC Genotyping Kit (Bayer HealthCare LLC, Tarrytown, NY), used in conjunction with COBAS AMPLICOR. Although the assay is sensitive, accurate, and easy to perform, additional optimization of the Invader 1.0 interpretive software (Invader Data Analysis Worksheet) may be necessary to reduce potential misidentification of HCV genotypes in low-titer specimens. In summary, Invader 1.0 is compatible with a variety of commercially available PCR-based HCV 5' NC region amplification assays and is suitable for routine HCV genotyping in clinical laboratories.

* Corresponding author. Mailing address: Division of Clinical Microbiology, Hilton 4-60, Mayo Clinic, 200 First Street S.W., Rochester, MN 55905. Phone: (507) 284-2255. Fax: (507) 284-4272. E-mail: jdcyao{at}mayo.edu.

Journal of Clinical Microbiology, February 2006, p. 318-323, Vol. 44, No. 2
0095-1137/06/$08.00+0     doi:10.1128/JCM.44.2.318-323.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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