Langaee Taimour, Stauffer Lynda, Galloway Cheryl, Solayman Mohamed H, Cavallari Larisa
Department of Pharmacotherapy and Translational Research, Center for Pharmacogenomics, University of Florida , College of Pharmacy, Gainesville, Florida.
Genet Test Mol Biomarkers. 2017 Apr;21(4):259-264. doi: 10.1089/gtmb.2016.0317. Epub 2017 Feb 21.
Developing genetic and pharmacogenetic panels enhances genetic testing in clinical molecular diagnostics and precision medicine. This study was designed to cross-validate the performance of Canon's multiplex high-resolution DNA melting analysis platform with the Applied Biosystems TaqMan-based Quant Studio Real-Time PCR System and Pyrosequencing genotyping platforms for common genetic polymorphisms of the vitamin K epoxide reductase complex 1 (VKORC1) and CYP2C9.
Genomic DNA isolated from 240 blood and saliva samples was used to genotype the VKORC1-1639 G/A (rs9923231), CYP2C92 (430C>T, rs28371674), and CYP2C93 (1075A>C, rs1057910) single-nucleotide polymorphisms (SNPs) on the three above-mentioned genotyping platforms.
There was 99.2%, 100%, and 100% concordance among the Canon DNA analyzer, the TaqMan-based QuantStudio, and the Pyrosequencing genotyping results for the VKORC1 (rs9923231), CYP2C92, and CYP2C93 SNPs, respectively, in DNA samples isolated from blood. The DNA samples isolated from saliva showed 100% concordance among the three test platforms for the three tested SNPs.
These results show that, the DNA analyzer performed very well when compared with two commonly used genotyping platforms. The reliability, multiple genetic variant testing capability, and short turnaround time for up to eight samples make the DNA analyzer an ideal genotyping platform for genetic testing in the clinical practice setting, where efficient genotyping is important to prevent delays in optimizing drug therapy.
开发基因和药物基因检测组合可增强临床分子诊断和精准医学中的基因检测。本研究旨在对佳能的多重高分辨率DNA熔解分析平台与应用生物系统公司基于TaqMan的Quant Studio实时荧光定量PCR系统以及焦磷酸测序基因分型平台进行性能交叉验证，以检测维生素K环氧化物还原酶复合体1（VKORC1）和CYP2C9的常见基因多态性。
从240份血液和唾液样本中分离出的基因组DNA用于在上述三种基因分型平台上对VKORC1 - 1639 G/A（rs9923231）、CYP2C92（430C>T，rs28371674）和CYP2C93（1075A>C，rs1057910）单核苷酸多态性（SNP）进行基因分型。
在从血液中分离的DNA样本中，佳能DNA分析仪、基于TaqMan的QuantStudio和焦磷酸测序基因分型结果对于VKORC1（rs9923231）、CYP2C92和CYP2C93 SNP的一致性分别为99.2%、100%和100%。从唾液中分离的DNA样本在三种检测平台对三种检测SNP的检测结果中显示出100%的一致性。
这些结果表明，与两种常用的基因分型平台相比，该DNA分析仪表现出色。其可靠性、多基因变异检测能力以及多达八个样本的短周转时间，使该DNA分析仪成为临床实践中基因检测的理想基因分型平台，在临床实践中高效基因分型对于防止优化药物治疗的延迟非常重要。
