摘要
目的建立一种用于检测亚甲基四氢叶酸还原酶(MTHFR)C667T基因型的方法,并对该方法进行评价。方法以MTHFR基因C667T位点为标靶设计引物,采用PCR-金磁微粒层析法结合扩增受阻突变体系PCR技术(ARMS-PCR)进行基因分型,将所建立的方法应用于临床样本检测。结果ARMS-PCR与金磁微粒层析法联合检测MTHFR C677T基因分型,全程耗时约60 min。检测最低限度为10^(3)copies/μL。PCR-金磁微粒层析法能特异性检测MTHFR基因分型,应用于50份人全血样本MTHFR基因分型检测,其结果与测序结果完全符合,符合率为100%(50/50)。结论PCR-金磁微粒层析法检测MTHFR C677T基因多态性方法具有特异性强、敏感性高、准确率高的特点,无需特殊大型设备,具有快速、便捷、准确、经济等优势,适合在各级医疗机构开展。
Objective To establish and evaluate a method for genotyping Methylenetetrahydrofolate reductase(MTHFR).Methods The MTHFR gene C667T site was used as the target to design primers.PCR gold magnetic particle chromatography combined with amplification refractory mutation system-PCR(ARMS-PCR)was used to detect the genotype.The established method was applied to clinical samples.Results The whole process of ARMS-PCR combined with PCR-gold magnetic particle chromatography could genotype MTHFR C667T in 60 minutes.The detection limit was 10^(3)copies/μL.PCR-gold magnetic particle chromatography can specifically detect MTHFR genotyping,and it was applied to detect MTHFR genotype using 50 human whole blood samples.The results were completely consistent with their sequencing results,with a coincidence rate of 100%(50/50).Conclusion The PCR-gold magnetic particle chromatography method for detecting MTHFR C677T gene polymorphism has the characteristics of strong specificity,high sensitivity,and high accuracy.It does not need special large equipment,and has the advantages of fast,convenient,accurate and economical,which is suitable for medical institutions at all levels.
作者
陈锦亮
蓝槐敏
黄慧莹
陈伟忠
CHEN Jinliang;LAN Huaimin;HUANG Huiying;CHEN Weizhong(Department of Medical Laboratory,Chaozhou People's Hospital Affiliated to Shantou University Medical College,Chaozhou,Guangdong,China,521011;Department of Medicament,Chaozhou People's Hospital Affiliated to Shantou University Medical College,Chaozhou,Guangdong,China,521011;Department of Medical Laboratory,Shenzhen Luohu Hospital Group,Shenzhen,Guangdong,China,518001)
出处
《分子诊断与治疗杂志》
2023年第12期2237-2241,共5页
Journal of Molecular Diagnostics and Therapy
基金
潮州市科技局项目(2020GY10)
潮州市卫生健康局项目(潮卫科研2020009)。
