摘要
目的：探讨通过扩增阻碍突变系统聚合酶链反应（ARMS—PCR）方法，在单个PCR反应体系内同时检测MTHFR677C〉T、1298A〉C和MTRR66A〉G基因多态性。方法：选取2012年7月～2013年3月在郑州大学第三附属医院（河南省妇幼保健院）进行孕前或孕期检查的育龄女性135名，留取空腹静脉血通过ARMS—PCR检测MTHFR677C〉T、1298A〉C和MTRR66A〉G基因多态性，同时随机留取30名研究对象口腔黏膜细胞，利用Taqman—MGB技术，通过荧光定量PCR方法验证ARMS—PCR结果。结果：ARMS—PCR产物电泳后条带清晰，易于判读3个位点SNP基因型；荧光定量PCR结果显示，30份样本中，两种方法的基因分型结果完全一致。郑州地区汉族育龄女性中，677T等位基因频率为66．3％，TT纯合子占42．9％；1298C等位基因频率为11．1％，纯合子CC占0．8％；66G等位基因频率为23．3％，纯合子GG占3．0％。结论：ARMS—PCR同时检测MTHFR677C〉T、1298A〉C和MTRR66A〉G基因多态性具有高效、经济、快速的特点，值得在SNP分型中使用。
Objective： To explore the methods which utilize 12 primers in a single PCR reaction to detect 677 C 〉 T and 1298 A 〉 C of Methylene tetrahydrofolate reductase （MTHFR） gene and 66 A 〉 G of the Methionine synthase reductase （MTRR） gene polymorphisms simultaneously based on the principle of amplification refractory mutation system PCR （ARMS - PCR） . Methods： Data collected from 135 women of childbearing age, whom pregnancy or unpregnancy for prenatal health care, from July 2012 to March 2013 at the Third Affiliated Hospital of Zhengzhou University. Fasting blood sample were collected to detect MTHFR C677T, A1298C and MTRR A66G polymorphisms by ARMS - PCR. In order to validate the ARMS - PCR genotyping results, at the same time thirty subjects mucous membrane of oral cavity were collected randomly for detection of the three polymorphisms by fluorescent quantization PCR separately, which based on the Taqman - MGB technique. Results： The bands of ARMS - PCR product agarose gel eleetrophoretogram were clear sufficiently to identify genotype. Furthermore genotype assignment was validated for thirty different samples by the fluorescent quantization PCR showed that there were no difference in genotype assignment was observed in any of the samples. The frequency of 677T allele was 66. 3% , TF homozygotes accounted for 42.9% ; 1298C allele frequency was 11.1%, homozygous CC accounting for 0. 8% ; 66G allele frequency was 23.3%, homozygous GG accounting for 3.0% among women of childbearing age in Zhengzhou. Conclusion： ARMS - PCR offering fast and economical simultaneous detection of MTHFR C677T and A1289C, and MTRR A66G polymorphisms and can be routinely used in SNP typing.
出处
《中国妇幼保健》
CAS
北大核心
2014年第8期1255-1257,共3页
Maternal and Child Health Care of China
基金
河南省卫生厅科技攻关项目〔201203050〕
河南省计划生育科学技术研究计划(2012)
河南省计划生育科学技术研究计划(2013)
