[1]尹栩芳,蒋卫霞,饶品彬.LNA-taqman探针实时荧光PCR快速检测CYP4F2-C1347T,CYP2C9*3,VKORC1-C1173T与VKORC1-G1639A基因多态性方法的建立[J].现代检验医学杂志,2017,32(06):1-5,10.[doi:10.3969/j.issn.1671-7414.2017.06.001]
　YIN Xu-fang,JIANG Wei-xia,RAO Pin-bin.Establishment of LNA-taqman Quantitative Real-time PCR for Detecting CYP4F2-C1347T,CYP2C9*3,VKORC1-C1173T and VKORC1-G1639A Genes Polymorphism[J].Journal of Modern Laboratory Medicine,2017,32(06):1-5,10.[doi:10.3969/j.issn.1671-7414.2017.06.001]
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LNA-taqman探针实时荧光PCR快速检测CYP4F2-C1347T,CYP2C9*3,VKORC1-C1173T与VKORC1-G1639A基因多态性方法的建立()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第32卷
期数:
2017年06期
页码:
1-5,10
栏目:
论著
出版日期:
2017-12-25

文章信息/Info

Title:
Establishment of LNA-taqman Quantitative Real-time PCR for Detecting CYP4F2-C1347T,CYP2C9*3,VKORC1-C1173T and VKORC1-G1639A Genes Polymorphism
文章编号:
1671-7414(2017)06-001-06
作者:
尹栩芳1; 蒋卫霞1; 饶品彬2
1.上海交通大学医学院附属苏州九龙医院检验科, 江苏苏州 215000; 2.苏州吉玛基因股份有限公司,江苏苏州 215000
Author(s):
YIN Xu-fang1; JIANG Wei-xia1; RAO Pin-bin2
1.Department of Clinical Laboratory, Suzhou Kowloon Hospital Affiliated to Shanghai Jiaotong University Schoolof Medicine, Jiangsu Suzhou 215000,China; 2.Suzhou Gene Pharma Co.,Ltd,Jiangsu Suzhou 215000,China
关键词:
华法林;  CYP4F2;  CYP2C9;  VKORC1
分类号:
Q503
DOI:
10.3969/j.issn.1671-7414.2017.06.001
文献标志码:
A
摘要:
目的 建立一种基于LNA-taqman探针实时荧光PCR检测华法林剂量相关基因型维生素K单氧酶CYP4F2,维生素K环氧化物还原酶复合体亚单位1(VKORC1)和细胞色素P4502C9(CYP2C9)的方法。方法 针对CYP4F2-1347C>T,CYP2C9*3,VKORC1-1173C>T与VKORC1-1639G>A四个单核苷酸多态性(SNP)位点,分别设计一套等位基因特异性探针与引物,并对LNA-taqman探针特异性进行评价。提取150例接受华法林治疗患者外周血样本基因组DNA进行检测,同时对部分实时荧光PCR产物样品进行验证。结果 ①LNA-taqman探针检测方法特异性高,没有交叉性。②150例患者中,CYP4F2-1347C>T基因型CC,CT和TT分别有87,56和7例,各占58%,37.3%和4.7%; CYP2C9*3基因型*1/*1和*1/*3分别有142和8例,各占94.7%和5.3%,未检出纯合子*3/*3基因型; VKORC1-1173C>T基因型TT,TC和CC分别有127,20和3例,各占84.7%,13.3%和2%; VKORC1-1639G>A基因型AA,AG和GG分别有124,23和3例,各占82.7%,15.3%和2%。结论 基于LNA-taqman探针的实时荧光PCR分型方法具有操作简便,价格低廉,结果准确可靠等特点,适于临床实验室对CYP4F2,CYP2C9*3,VKORC1的基因分型。
Abstract:
Abstract:Objective To establish a LNA-taqman-based real-time PCR assay for detecting gene polymorphisms of Vitamin K monooxygenase CYP4F2,vitamin K epoxide reductase complex subunit 1(VKORC1)and cytochrome P4502C9(CYP2C9),associated with Warfarin optimal dosage.Methods A set of allele-specific PCR primers and probes was designed for each single nucleotide polymorphism(SNP)of CYP4F2-1347C>T,CYP2C9*3,VKORC1-1173C>Tand VKORC1-1639G>A,and the specificity of LNA-taqman probe PCR was evaluated.Genomic DNA of peripheral blood samples from 150 patients with treated with warfarin was extracted,and the some PCR products were verified with sequencing.Results ①LNA-taqman-based real-time PCR assay was highly specificity,no overla.②Among the 150 patients,the cases of CC,CT and TTgenotype of CYP4F2-1347C>T were 87(58%),56(37.3%)and 7(4.7%).The cases of*1/*1 and *1/*3 genotype of CYP2C9*3 were 142(94.7%)and 8(5.3%),*3/*3 genotype was not detected.The cases of TT,TC and CC genotype of VKORC1 1173C>T were127(84.7%),20(13.3%)and 3(2%).The cases of AA,AG and GG genotype of VKORC1 1639G>A were 124(82.7%),23(15.3%)and 3(2%),respectively.Conclusion The LNA-taqman-based real-time PCR assay is convenient,inexpensive,accurate and will be useful for CYP4F2-C1347T,CYP2C9*3,VKORC1-C1173T and VKORC1-G1639A genotyping in a clinic laboratory.

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备注/Memo

备注/Memo:
作者简介:尹栩芳(1974-),女,本科,副主任技师,主要从事临床生物化学、免疫学检验研究,E-mail:yxfd1225@sina.com。 通讯作者:饶品彬(1989-),男,硕士,苏州吉玛基因股份有限公司,主要从事基因检测研究,E-mail:pinbinrao@163.com。
更新日期/Last Update: 2017-12-26