飞行时间质谱与荧光定量法检测叶酸代谢基因多态性的比较  

Comparison of MALDI TOF MS and fluorescent quantitation for folate metabolism enzymes SNPs detection

作　　者：常亮[1] 任妲妮 焦利萍[1] 刘娟 李乾 刘平[1] CHANG Liang;REN Dani;JIAO Liping;LIU Juan;LI Qian;LIU Ping(Center for Reproductive Medicine, Department of Obstetrics and Gynecology, Peking University Third Hospital,National Clinical Research Center for Obstetrics and Gynecology(Peking University Third Hospital),Key Laboratory of Assisted Reproduction (Peking University), Ministry of Education Beijing Key Laboratory of Reproductive Endocrinology and Assisted Reproductive Technology, Beijing, 100191;Institute of Science and Technology, National Health Commission)

机构地区：[1]北京大学第三医院妇产科生殖医学中心,国家妇产疾病临床医学研究中(北京大学第三医院),辅助生殖教育部重点实验室(北京大学),北京市生殖内分泌与辅助生殖技术重点实验室,100191 [2]国家卫生健康委科学技术研究所

出　　处：《中国计划生育学杂志》2021年第7期1411-1413,1417,1541,共5页Chinese Journal of Family Planning

基　　金：北京市自然科学基金面上项目(编号7202226);国家重大研发计划(编号2018YFC1003100)。

摘　　要：目的:评价基质辅助激光解析电离飞行时间质谱(MALDI-TOF MS)技术用于叶酸代谢关键酶基因临床检测的潜力。方法:以在本院就诊不孕症患者的100份EDTA抗凝静脉全血为样本,应用MALDI-TOF MS检测体系检测叶酸代谢关键酶基因的单核苷酸多态性(SNP)位点,包括5,10-亚甲基四氢叶酸还原酶(MTHFR)基因2个多态性位点677 C>T和1298 A>C,以及甲硫氨酸合成酶还原酶(MTRR)基因1个多态性位点66 A>G。将检测结果与实时荧光定量聚合酶链式反应(RT-qPCR)检测MTHFR 677 C>T和1298 A>C的结果进行对比。结果:MALDI-TOF MS与RT-qPCR均检出677 C>T位点CC型21例,CT型43例,TT型36例;1298 A>C位点AA型78例,AC型20例,CC型2例,二者检测结果的符合率为100%;MALDI-TOF MS系统还同时检测MTRR 66 A>G位点AA型50例,AG型44例,GG型6例。结论:MALDI-TOF MS检测体系可应用于叶酸代谢关键酶基因多态性的检测,与RT-qPCR相比具有通量高、准确度高的特点。Objective:To evaluate the application value of the matrix assisted laser desorption ionization time of flight mass spectrometry(MALDI TOF MS)for folate metabolism enzymes SNPs detection.Methods:A total of 100 women diagnosed with infertility were included.EDTA anticoagulant peripheral blood of these women were used for detecting single nucleotide polymorphisms(SNPs)of key regulatory enzymes involved in folate metabolism,including C677T and A1298C of 5,10 methylenetetrahydrofolate reductase(MTHFR)and A66G of methionine synthase reductase(MTRR)by MALDI TOF MS.In addition,the consistence of MALDI TOF MS with real time fluorescence quantitative polymerase chain reaction(RT qPCR)for detection was compared.Results:MALDI TOF MS and RT qPCR had detected 21 cases of CC,43 cases of CT,and 36 cases of TT on MTHFR 677 C>T,and 78 cases of AA,20 cases of AC,and 2 cases of CC on 1298 A>C.The coincidence rates of the two methods were all 100%,which had no significant differences for genotype distribution of MTHFR C677T and A1298C between two the methods.In the same time,MALDI TOF MS was used to detect 50 cases of AA on MTRR 66 A>G,44 cases of AG on MTRR 66 A>G,and 6 cases of GG on MTRR 66 A>G.Conclusion:MALDI TOF MS can be applied to detecting the key regulatory enzymes SNPs with the advantage of high throughput and accuracy compared with RT qPCR.

关 键 词：基质辅助激光解吸电离飞行时间质谱 实时荧光定量聚合酶链式反应 5 10-亚甲基四氢叶酸还原酶 甲硫氨酸合成酶还原酶 

分 类 号：R73[医药卫生—肿瘤]