RESUMEN
Presently, clopidogrel is the standard therapeutic drugs for antiplatelet therapy. Variants in the CYP2C19 gene influence the clinical response of clopidogrel. Thus, the US Food and Drug Administration suggested CYP2C19 genotyping needs to identify before taking medicine. Due to high cost, time consuming, and sophisticated instruments, current single nucleotide polymorphism detection methods are limited in clinical application. In the present study, we established a genotyping method for CYP2C19, which combines amplification refractory mutation system (ARMS)-PCR with a lateral flow assay used gold magnetic nanoparticles (GMNPs) named as PCR-gold magnetic lateral flow assay system (PCR-GoldMag LFA). The PCR products with specific genotype can be explained within 5 minutes, either through visual or by a magnetic reader automatically according to the captured GMNPs probes on the test lines of the LFA strips. The limit of detection of this method is 5 ng of genomic DNA. The PCR-GoldMag LFA system was applied in a clinical trial with 1356 samples of Han Chinese. The concordance rate between the LFA system and sequencing is 99.93%. The allele frequency of CYP2C19*2 and CYP2C19*3 are 30.38 and 7.08% in Han Chinese, respectively. This method provides a new way in the clinical application of CYP2C19 genotyping to guide the clopidogrel medication.
Asunto(s)
Citocromo P-450 CYP2C19/genética , Técnicas de Genotipaje/instrumentación , Oro/química , Nanopartículas de Magnetita/química , Reacción en Cadena de la Polimerasa , Citocromo P-450 CYP2C19/sangre , Frecuencia de los Genes , Genotipo , Técnicas de Genotipaje/economía , Humanos , Mutación , Reacción en Cadena de la Polimerasa/economía , Sensibilidad y Especificidad , Análisis de Secuencia de ADN/normas , Factores de TiempoRESUMEN
Objective To estabolish the ALDH2 Glu504Lys genotyping technique for oral swab sample type based on PCR-gold magnetic nanoparticle chromatographyl.Methods According to the ALDH2 Glu504Lys gene the specific primers were designed by ARMS-PCR technique.Optimized the optimal ALDH2 Glu504Lys oral swab reaction system and PCR reaction procedures based on the routine reaction conditions and reaction procedure of laboratory PCR.60 samples of oral swabs were collected and tested by gold magnetic nano-chromatography.The results were verified by sequencing.Results The detected 60 cases of oral swab samples contain 16 cases heterozygous mutation,wild type 42 cases,homozygous mutation 2 cases and then the results by Goldmag nanoparticle detection was perfectly matched with sequencing results.Conclusion The Goldmag nanoparticles chromatographic detection technique of ALDH2 Glu504Lys buccal swabs system was established.With accuracy,fastness,reliability,it is worthy of clinical promotion.