Clinical application and importance of one-step human CYP2C19 genotype detection

Abstract

BackgroundTo directly achieve cytochrome P450 2C19 gene (CYP2C19) classification using one-step real-time fluorescent PCR detection and to verify the capabilities of this method with nucleic acid extracted from whole blood samples.MethodsA human CYP2C19 genotyping kit based on one-step real-time fluorescent PCR detection was used to analyze whole blood or genomic DNA samples. This method was compared with pyrosequencing and another quantitative (q)PCR kit for its accuracy, repeatability, detection range analysis, sensitivity, specificity, and anti-interference analysis.ResultsThe one-step real-time PCR method achieved a 100% accuracy rate compared with pyrosequencing and the other qPCR kit. When detecting different concentrations of known genes, concentrations of each sample ranging from 0.2 to 125 ng/µL could be correctly detected. The genotypes of samples treated with anticoagulants, including EDTA and sodium citrate, and chyle blood samples could be correctly detected.ConclusionThe one-step detection method demonstrated high accuracy and a wide detection range. It also had high levels of repeatability, sensitivity, and specificity for the assessment of genomic DNA test samples.

Molecular Genetic Techniques in Biomarker Analysis Relevant for Drugs Centrally Approved in Europe

“…However, for genotyping PCR can also directly be used for detection and is a cost-effective method [ 13 ]. Furthermore, techniques such as real-time PCR (qPCR) [ 14 ] and real-time quantitative RT-PCR (RT-qPCR) [ 15 ], the amplification refractory mutation system (ARMS) [ 16 ] or multiplex ligation-dependent probe amplification (MLPA) [ 17 ] are also used as primary mutation or variant-detection systems. Nevertheless, they can only detect mutations that have been previously characterized and thus are suitable for a targeted analysis [ 18 ].…”

Analysis The Effect of Gene and Platelet Testing Guide Dual Anti-Upgrade Therapy After PCI in Patients With ACS

“…The current gene detection methods of individualized medicine [4] mainly include: PCR direct sequencing method, PCR pyrophosphate sequencing method, quantitative uorescence PCR method, PCR gene chip method, PCR electrophoresis analysis method, high resolution melting analysis method, allele polymorphism, in situ hybridization, etc. Among them, realtime uorescence PCR is the most popular because of its high sensitivity, accurate classi cation and convenient operation.…”

“…the system is a fully automated genome extraction, preparation and analysis system with an analysis time of about 2 to 4 hours. There is also a portable system with low entry cost STQ3 [17], which contains a one-step method CYP2C19 genotyping detection kit (Coyote Bioscience) [4]. compared with other kits, direct use of whole blood without nucleic acid extraction reduces test time and cost.…”

“…Patients after Percutaneous coronary intervention(PCI) surgery often require dual antiplatelet therapy Double antiplatelet therapy(DAPT) for at least 12 months [1], usually aspirin and P2Y12 receptor inhibitors such as clopidogrel, prasugrel, or ticagrelor [2]. However, factors such as environmental, clinical, and genetic variation can in uence differences in antiplatelet responses and increase the risk of thromboembolism events, the stent, especially in clopidogrel [3], but not in prasugrel or ticagrelor [4]. How to select P2Y12 inhibitors, in addition to the clinical assessment of the risk of ischemia/bleeding, gene and platelet function testing has gradually provided the basis for individualized accurate treatment and has been paid more and more attention [5].…”