Evaluation of the Verigene Warfarin Metabolism Nucleic Acid Test Kit for the Rapid Detection of CYP2C9 and VKORC1 Polymorphisms

Hyeyoung Lee; Jiyeon Kim; Gun Dong Lee; Joonhong Park; Hyojin Chae; Myungshin Kim; Yonggoo Kim; Yong Seog Oh

doi:10.3343/lmo.2014.4.4.198

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Lee, Kim, Lee, Park, Chae, Kim, Kim, and Oh: Evaluation of the Verigene Warfarin Metabolism Nucleic Acid Test Kit for the Rapid Detection of CYP2C9 and VKORC1 Polymorphisms

Original Article

Diagnostic Genetics

Laboratory Medicine Online 2014; 4(4): 198-202.

Published online: 1 October 2014

DOI: https://doi.org/10.3343/lmo.2014.4.4.198

Evaluation of the Verigene Warfarin Metabolism Nucleic Acid Test Kit for the Rapid Detection of CYP2C9 and VKORC1 Polymorphisms

Hyeyoung Lee, M.D.¹, Jiyeon Kim, M.T.², Gun Dong Lee, M.T.¹, Joonhong Park, M.D.^1,², Hyojin Chae, M.D.^1,², Myungshin Kim, M.D.^1,², Yonggoo Kim, M.D.^1,², Yong Seog Oh, M.D.³

¹Department of Laboratory Medicine, College of Medicine, The Catholic University of Korea, Seoul, Korea.

²Catholic Laboratory Development and Evaluation Center, College of Medicine, The Catholic University of Korea, Seoul, Korea.

³Department of Internal Medicine, Division of Cardiology, Seoul St. Mary's Hospital, Seoul, Korea.

Corresponding author: Hyojin Chae. Department of Laboratory Medicine, Seoul St. Mary's Hospital, College of Medicine, The Catholic University of Korea, 222 Banpodae-ro, Seocho-gu, Seoul 137-701, Korea. Tel: +82-2-2258-1646, Fax: +82-2-2258-1719, chez@catholic.ac.kr

Received 8 March 2013 Revised 29 January 2014 Accepted 13 March 2014

(open-access):

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Background

Warfarin is a widely used oral agent for anticoagulation therapy. Warfarin has a narrow therapeutic index and a wide variation in the interindividual therapeutic dosage. Recently, genotypes of CYP2C9 and VKORC1 have been found to account for 30-40% of the warfarin dosing variability, and a variety of commercial genotyping assays are being introduced. In this study, we evaluated the Verigene Warfarin Metabolism Nucleic Acid test (Verigene Warfarin assay; Nanosphere, USA) for its accuracy and clinical utility in genotyping CYP2C9*2, CYP2C9*3, and VKORC1 1173C>T.

Methods

We compared the Verigene Warfarin assay with direct sequencing for accuracy in determining the genotypes of CYP2C9*2, CYP2C9*3, and VKORC1 1173C>T using 50 patient samples and 3 commercial DNA samples with known genotypes. The method was also evaluated for turn-around time, hands-on time, and feasibility.

Results

The Verigene Warfarin assay demonstrated 100% accuracy for identifying CYP2C9*2, CYP2C9*3, and VKORC1 1173C>T. The turn-around time and hands-on time were 3 hr and 2 min, respectively. The no-call error rate at first attempt was estimated to be 2%.

Conclusions

The Verigene Warfarin assay provides rapid and accurate genotype results. Considering there are only a few steps requiring manual intervention, it would be feasible to implement this assay even in clinical laboratories that lack considerable expertise in molecular diagnostics.

Keywords: Warfarin, CYP2C9, VKORC1

Figures and Tables

Fig. 1

Sequencing chromatograms for the respective genotypes of (A) CYP2C9*2 (wild type homozygous, mutant heterozygous, mutant homozygous), (B) CYP2C9*3 (wild type homozygous, mutant heterozygous), and (C) VKORC1 (wild type homozygous, mutant heterozygous, mutant homozygous).

Table 1

PCR primer sequences for CYP2C9*2, CYP2C9*3, and VKORC1 (1173C>T)

Abbreviations: Tm, melting temperature; bp, base pair.

Table 2

Comparison of results of Verigene Warfarin Metabolism Nucleic Acid Test and direct sequencing for CYP2C9*2, CYP2C9*3, and VKORC1

Notes

This article is available from http://www.labmedonline.org

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