荧光PCR探针熔解曲线法检测珠海市孕妇MTHFR和MTRR基因多态性研究  

Research of the fluorescence PCR based probe melting curve analysis in the detection of MTHFR and MTRR gene polymorphism among Zhuhai pregnancy

作　　者：李恋湘 肖奇志[1] 王格[1] 周玉球[1] 李磊[1] 胡玲玲[1] LI Lianxiang;XIAO Qizhi;WANG Ge;ZHOU Yuqiu;LI Lei;HU Lingling(Zhuhai Municipal Maternity and Child Healthcare Hospital,Zhuhai Institute of Medical Genetics&Department of Clinical Laboratory,Zhuhai 519001,China)

机构地区：[1]珠海市妇幼保健院,珠海市医学遗传研究所,广东珠海519001

出　　处：《临床医药实践》2020年第8期608-612,640,共6页Proceeding of Clinical Medicine

摘　　要：目的:评价荧光PCR探针熔解曲线法(PMCA)检测亚甲基四氢叶酸还原酶(MTHFR)C677T,A1298C和蛋氨酸合成酶还原酶(MTRR)A66G基因多态性的适用性,并探讨珠海市孕妇各基因多态性分布特点及同型半胱氨酸(Hcy)水平情况。方法:通过双盲方式,对珠海市400例孕妇分别采用PMCA技术和DNA测序技术检测MTHFR C677T,A1298C和MTRR A66G基因多态性,以DNA测序为标准,评价PMCA法的检测效能。统计各检测位点的基因型频率,对孕妇叶酸利用能力的遗传风险进行评估,并分析其与血清同型半胱氨酸水平的相关性。结果:PMCA法检测上述基因多态性的结果与DNA测序结果完全吻合。珠海市孕妇MTHFR C677T位点TT,MTHFR A1298C位点CC,MTRR A66G位点GG纯合子频率分别为12.5%,1.7%和6.8%;MTRR A66G位点AG杂合子频率34.0%。高风险组血清Hcy水平与未发现风险组比较差异有统计学意义(t=-4.958,P<0.05)。结论:PMCA技术能快速准确检测MTHFR和MTRR基因多态性。珠海市孕妇MTHFR和MTRR基因型频率分布有其自身特点。孕妇血清同型半胱氨酸水平对临床叶酸个性化用药有指导作用。Objective:To evaluate the clinical applicability of the fluorescence PCR based probe melting curve analysis(PMCA)in the detection of MTHFR C677 T,A1298 C and MTRR A66 G gene polymorphism,and to analyze the distribution characteristics of gene polymorphism and homocysteine levels among Zhuhai pregnancy.Methods:The PMCA assay and the direct DNA sequencing analysis were respectively used to detect MTHFR C677 T,A1298 C and MTRR A66 G gene polymorphisms in a double-blind way,and the detection performance of the PMCA assay was evaluated through the standard of direct DNA sequencing analysis;In addition,we also analyzed the relativity between the genetic risk of folic acid utilization ability and the homocysteine levels by using the statistics such as gene frequency of each detection point.Results:The results of the PMCA assay and the direct DNA sequencing analysis were completely the same.The frequencies of MTHFR C677 T TT,MTHFR A1298 C CC and MTRR A66 G GG homozygous genotype were 12.5%,1.7% and 6.8% respectively,and the frequency of MTRR A66 G AG heterozygous genotype was 34.0% among Zhuhai pregnancy.There was a statistically significant difference in homocysteine values between the group with genetic risk not found yet and the group of high genetic risk(t=-4.958,P<0.05).Conclusions:The PMCA assay can be used in fast and accurate detection of MTHFR and MTRR gene polymorphisms,the gene frequency of which shows their own distribution characteristics among Zhuhai pregnancy.The homocysteine levels of the pregnancy can be used as a guide in clinical personalized supplementation of folic acid.

关 键 词：熔解曲线 叶酸 亚甲基四氢叶酸还原酶 蛋氨酸合成酶还原酶 

分 类 号：R722.1[医药卫生—儿科]