MALDI-TOF-MS和RT-qPCR对于SLCO1B1和ApoE多态性检测的比较  

Comparison of MALDI-TOF-MS and RT-qPCR for detection of SLCO1B1 and ApoE gene polymorphisms

作　　者：叶阿里[1] 邹雨桐 张海燕 吴洁[1] 张睿[1] 张晓峰 马庆伟 YE A-li;ZOU Yu-tong;ZHANG Hai-yan;WU Jie;ZHANG Rui;ZHANG Xiao-feng;MA Qing-wei(Department of Clinical Laboratory,Peking Union Medical College Hospital,CAMS&PUMC,Beijing 100730;Bioyong(Beijing)Technologies Inc.Ltd.Beijing 102206,China)

机构地区：[1]中国医学科学院,北京协和医学院,北京协和医院检验科,北京100730 [2]北京毅新博创生物科技有限公司,北京102206

出　　处：《基础医学与临床》2020年第7期980-985,共6页Basic and Clinical Medicine

基　　金：国家自然科学基金青年科学基金(81301508)。

摘　　要：目的探讨基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS)和实时荧光定量PCR(RT-qPCR)检测脂质及药物代谢相关的基因位点的方法学比对,以评估MALDI-TOF-MS是否可用于临床SLCO1B1和ApoE多态性检测。方法纳入2018年7月至9月在北京协和医院诊断为高脂血症、高血压或动脉粥样硬化的患者共计71例。收集EDTA抗凝全血并提取基因组DNA,分别采用MALDI-TOF-MS和RT-qPCR检测SLCO1B1的rs2306283(388A>G)和rs4149056(521T>C)位点以及ApoE的rs429358(388T>C)和rs7412(526C>T)位点,并利用Kappa系数比较结果的一致性。结果两种方法对相关基因位点的检测结果具有较高的一致性,仅1例患者rs4149056(521T>C)位点出现分型差异且在MALDI-TOF-MS复检后得到与RT-qPCR一致的结果。结论MALDI-TOF-MS可应用于脂质及药物代谢相关基因SLCO1B1和ApoE多态性的检测,从而指导临床他汀类用药。Objective To compare the consistence between matrix-assisted laser desorption ionization time of flight mass spectrometry(MALDI-TOF-MS)with real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)on genetic polymorphisms associated with the metabolism of lipids or drugs,and assess whether the MALDI-TOF-MS method can be used for clinical SLCO1B1 and ApoE gene polymorphism detection.Methods A total of 71 individuals diagnosed with hyperlipidemia,hypertension or atherosclerosis in Peking Union Medical College Hospital were included.The whole blood samples with EDTA were collected and genomic DNA was extracted.MALDI-TOF-MS and RT-qPCR were applied for analyzing rs2306283(388A>G)and rs4149056(521T>C)of SLCO1B1 as well as rs429358(388T>C)and rs7412(526C>T)of ApoE.The results of genetic polymorphisms were compared by Kappa coefficient.Results There were no statistically significant differences for genotype distribution of SLCO1B1 and ApoE between two methods.Only one sample had a typing difference in the rs4149056(521T>C)locus between two methods.However,the results consistent with RT-qPCR were obtained after the sample was retested by MALDI-TOF-MS.Conclusions MALDI-TOF-MS can be applied to the detection of lipid and drug metabolism-related genes SLCO1B1 and ApoE polymorphisms,thereby guiding statins treatment.

关 键 词：基质辅助激光解析电离飞行时间质谱 实时荧光定量PCR 基因多态性 他汀类药物 

分 类 号：R34[医药卫生—基础医学]