生殖医学杂志2024，Vol.33Issue(6)：785-791,7.DOI:10.3969/j.issn.1004-3845.2024.06.013
基于CRISPR/Cas12a的叶酸代谢位点MTHFR基因C677T多态性检测方法的建立
Establishment of adetection method for MTHFR gene C677T polymorphism in folate metabolism using CRISPR/Cas12a
摘要
Abstract
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.关键词
人亚甲基四氢叶酸还原酶基因/多态性/CRISPR/Cas12a/恒温扩增Key words
Human methylene tetrahydrofolate reductase gene/Polymorphism/CRISPR/Cas12a/Isothermal amplification分类
医药卫生引用本文复制引用
田石,安莉莎,马瑶,金孝华,马旭,张璐..基于CRISPR/Cas12a的叶酸代谢位点MTHFR基因C677T多态性检测方法的建立[J].生殖医学杂志,2024,33(6):785-791,7.基金项目
国家自然科学基金(82200261) （82200261）
海淀区属卫生健康系统高层次人才发展计划项目(2022HDXD006) （2022HDXD006）
