An extraction‐free method for rapid detection of CYP2C19 * 2/3/17 polymorphisms in one tube using melting curve analysis

• DOI: 10.1002/biot.202300207
• Publication Date: 2023-08-08T11:09:18Z
• Authors: Jianguang Guo; Weixin You; Kangfeng Lin; Qinghan Li; Xiangju Guo; Shuai Wang; Ya Bian; Wenjing Ren; Rui Zhang; Yanping Wang; Boan Li
• Keywords: Melting curve analysis; High Resolution Melt; Sanger sequencing

Drug-metabolizing enzymes play an important role in the metabolism of drugs vivo. Their activity is factor affecting rate drug metabolism, which directly determines intensity and persistence action. Patients taking medication can be divided into different metabolic types through detection CYP2C19 drug-metabolizing enzyme gene polymorphisms, then used for guidance clopidogrel. Here, we describe a method based on real-time polymerase chain reaction (PCR). This uses multicolor melting curve analysis to accurately identify mutation sites genotypes * 2, 3, 17. The limit plasmid samples was 1 copies μL-1 ; that genomic 0.1 ng . system detect nine 2/3/17 at three one tube, quickly achieving within h. Combined with sample release agent, extraction completed 5 s, rapid diagnosis without timely treatment. Furthermore, not limited blood also applied oropharyngeal saliva samples, increasing sampling diversity convenience. When using clinical (n = 93), established able genotypes, accuracy effectiveness were confirmed by Sanger sequencing. Due its accuracy, rapidity, simple operation, low cost, technology amplification combined expected become powerful tool detecting guiding clopidogrel use countries resources.