TY - JOUR
T1 - Development and Cross-Validation of High-Resolution Melting Analysis-Based Cardiovascular Pharmacogenetics Genotyping Panel
AU - Langaee, Taimour
AU - El Rouby, Nihal
AU - Stauffer, Lynda
AU - Galloway, Cheryl
AU - Cavallari, Larisa H.
N1 - Publisher Copyright:
© 2019 Mary Ann Liebert, Inc.
PY - 2019/3
Y1 - 2019/3
N2 - Aims: This study was designed to develop a high-resolution melting (HRM) analysis-based cardiovascular (CV) pharmacogenetics (PGx) genotyping panel for the Canon DNA Genetic Analyzer multiplex genotyping platform and cross-validate its performance with the TaqMan®-based OpenArray® method. Methods: The CV PGx genotyping panel containing 17 single nucleotide polymorphisms (SNPs) selected from 5 genes (CYP2C9, CYP2C19, CYP4F2, SLCO1B1, and VKORC1) and the CYP2C cluster was used to compare genotyping results between analysis methods. Genomic DNA from 223 clinical samples was used to genotype the 17 SNPs on the Canon DNA Genetic Analyzer and TaqMan OpenArray Quant Studio Real-Time PCR (polymerase chain reaction) System. Results: The concordance between the Canon DNA analyzer and TaqMan-based OpenArray genotyping results for the 17 SNPs ranged from 99.10% to 100% where SNPs (rs4244285, rs12248560, rs4986893, rs72552267, rs28399504, rs4149056, rs28371686, rs9332131, rs72558189, rs9923231, rs12777823), (rs41291556, rs1799853, rs7900194, rs28371685, rs2108622), and (rs1057910) showed 100%, 99.60%, and 99.10% concordance, respectively. Conclusion: These results show that the HRM analysis-based CV PGx genotyping panel performed well when compared with TaqMan-based OpenArray. The multiple genetic variant testing capability, efficient turnaround time and reproducibility of both assays formats suggest that the PGx panel with the DNA analyzer or other real-time PCR instruments with HRM assay analysis capability can be used for PGx testing in both research and clinical practice settings.
AB - Aims: This study was designed to develop a high-resolution melting (HRM) analysis-based cardiovascular (CV) pharmacogenetics (PGx) genotyping panel for the Canon DNA Genetic Analyzer multiplex genotyping platform and cross-validate its performance with the TaqMan®-based OpenArray® method. Methods: The CV PGx genotyping panel containing 17 single nucleotide polymorphisms (SNPs) selected from 5 genes (CYP2C9, CYP2C19, CYP4F2, SLCO1B1, and VKORC1) and the CYP2C cluster was used to compare genotyping results between analysis methods. Genomic DNA from 223 clinical samples was used to genotype the 17 SNPs on the Canon DNA Genetic Analyzer and TaqMan OpenArray Quant Studio Real-Time PCR (polymerase chain reaction) System. Results: The concordance between the Canon DNA analyzer and TaqMan-based OpenArray genotyping results for the 17 SNPs ranged from 99.10% to 100% where SNPs (rs4244285, rs12248560, rs4986893, rs72552267, rs28399504, rs4149056, rs28371686, rs9332131, rs72558189, rs9923231, rs12777823), (rs41291556, rs1799853, rs7900194, rs28371685, rs2108622), and (rs1057910) showed 100%, 99.60%, and 99.10% concordance, respectively. Conclusion: These results show that the HRM analysis-based CV PGx genotyping panel performed well when compared with TaqMan-based OpenArray. The multiple genetic variant testing capability, efficient turnaround time and reproducibility of both assays formats suggest that the PGx panel with the DNA analyzer or other real-time PCR instruments with HRM assay analysis capability can be used for PGx testing in both research and clinical practice settings.
KW - Cardiovascular pharmacogenetics genotyping panel
KW - high-resolution melting analysis
KW - microfluidic
KW - rapid serial PCR
UR - http://www.scopus.com/inward/record.url?scp=85062988564&partnerID=8YFLogxK
U2 - 10.1089/gtmb.2018.0298
DO - 10.1089/gtmb.2018.0298
M3 - Article
C2 - 30758238
AN - SCOPUS:85062988564
SN - 1945-0265
VL - 23
SP - 209
EP - 214
JO - Genetic Testing and Molecular Biomarkers
JF - Genetic Testing and Molecular Biomarkers
IS - 3
ER -