Establishment of adetection method for MTHFR gene C677T polymorphism in folate metabolism using CRISPR/Cas12a
Objective:To develop an efficient detection method based on CRISPR/Cas12a technology for the folate metabolism site c.677(MTHFR C677T),enabling rapid and accurate analysis of the C677T polymorphism.Methods:A single-tube detection system was established by combining recombinase polymerase amplification(RPA)with CRISPR/Cas12a to develop a genotyping strategy for the human MTHFR gene C677T polymorphism.Two hundred pregnant woman samples were tested for MTHFR gene C677T polymorphism,and the results were compared for concordance with conventional polymerase chain reaction(PCR)product sequencing.Results:The detection method for the human MTHFR gene C677T polymorphism,utilizing the CRISPR/Cas12a system,demonstrated high accuracy and excellent specificity.The results exhibited a remarkably high concordance with those obtained from PCR product sequencing.Conclusions:The established genotyping method for the human MTHFR C677T gene,utilizing CRISPR/Cas12a detection technology,is simple,rapid,and precise.This method provides a novel approach for the genotyping of the folate metabolism site c.677,holding promising potential for clinical applications.
Human methylene tetrahydrofolate reductase genePolymorphismCRISPR/Cas12aIsothermal amplification
国家科技期刊平台
NSTL
万方数据
