CAJ | 학술논문

目的：通过扩增阻滞突变系统-聚合酶链反应（ ARMS-PCR）检测H型高血压患者5，10-亚甲基四氢叶酸还原酶（MTHFR）C677T基因型，以期建立一种低成本、简单快速的MTHFR C677T基因分型方法。方法基于湖南省H型高血压危险因素研究的大型流行病调查的人群基础，采用单纯随机方法，选取94例H型高血压患者作为研究对象。设计阻滞度递减的系列错配引物，采用ARMS-PCR对94例患者外周血样本的MTHFR C677T基因型进行鉴定，并抽取不同基因型进行测序验证。结果 ARMS-PCR产物电泳后条带清晰，易于判读MTHFR C677T基因型。94例H型高血压患者共检出CC基因型42例、CT基因型24例和TT基因型28例。ARMS-PCR与DNA测序结果一致。结论 ARMS-PCR经阻滞度递减的系列错配引物能有效对MTHFR C677T基因型进行鉴定，此法具有成本低、操作简单、用时短的优点。适用于各类基因的多态性分析。
목적：통과확증조체돌변계통-취합매련반응（ ARMS-PCR）검측H형고혈압환자5，10-아갑기사경협산환원매（MTHFR）C677T기인형，이기건립일충저성본、간단쾌속적MTHFR C677T기인분형방법。방법기우호남성H형고혈압위험인소연구적대형류행병조사적인군기출，채용단순수궤방법，선취94례H형고혈압환자작위연구대상。설계조체도체감적계렬착배인물，채용ARMS-PCR대94례환자외주혈양본적MTHFR C677T기인형진행감정，병추취불동기인형진행측서험증。결과 ARMS-PCR산물전영후조대청석，역우판독MTHFR C677T기인형。94례H형고혈압환자공검출CC기인형42례、CT기인형24례화TT기인형28례。ARMS-PCR여DNA측서결과일치。결론 ARMS-PCR경조체도체감적계렬착배인물능유효대MTHFR C677T기인형진행감정，차법구유성본저、조작간단、용시단적우점。괄용우각류기인적다태성분석。
Objective To develop a cost -effective,rapid and simple method for MTHFR C677T genotyping by amplification refractory mutation system - polymerase chain reaction ( ARMS - PCR ) on H - type hypertension patients. Methods From the population sampled by the large epidemiological investigation based on the research of risk factors for H-type hypertension in Hunan Province,94 H-type hypertension patients were taken as subjects using simple random method. A series of mismatched -primers of refractory reduced were designed,and ARMS -PCR was employed to identify the MTHFR C677T genotypes of peripheral blood samples of the 94 patients, and different genotypes were abstracted for sequencing validation. Results The bands of ARMS - PCR product agarose gel electrophoretogram were clear sufficiently to identify genotype. Among the 94 H-type hypertension patients 42 were CC genotype,24 were CT genotype and 28 were TT genotype. ARMS-PCR results were in complete accord with DNA sequencing. Conclusion ARMS -PCR is a low -cost,simple and time- saving method for the determination of MTHFR C677T genotype by mismatched - primers of refractory reduced. The approach can be used to detect other gene polymorphism.