摘要
目的对华法林代谢相关的人细胞色素P450超家族第二亚家族C9代谢酶(CYP2C9)和维生素K环氧化物还原酶复合体亚单位1(VKORC1)基因多态性检测试剂盒进行性能验证。方法根据中国合格评定国家认可委员会(CNAS)制定的有关要求,结合试剂说明书,从符合率、精密度、检出限、抗干扰能力4个方面对苏州旷远生物分子技术有限公司的人CYP2C9与VKORC1基因多态性检测试剂盒(荧光PCR法)进行性能验证。结果20例临床标本的荧光PCR检测结果与Sanger测序法检测结果的符合率为100%;荧光PCR对1例临床标本重复检测15次,CYP2C9与VKORC1基因荧光通道的Ct值变异系数(CV)均<5%;试剂盒最低检测限为10 ng/μL;已知基因型的血液标本中加入干扰物血红蛋白、胆红素和胆固醇之后的基因型结果与对照组完全一致。结论人CYP2C9与VKORC1基因多态性检测试剂盒(荧光PCR法)符合ISO15189质量管理要求,可以在保证项目验收的基础上,为临床提供可靠的检测结果。
Objective To verify the performance characteristics of warfarin metabolism-related human CYP2C9 and VKORC1 gene polymorphisms detection kit(quantitative PCR).Methods The verification protocol designed by China National Accreditation(CNAS)combined with reagent manual,performance of the human CYP2C9 and VKORC1 gene polymorphisms detection kit manufactured by Suzhou Kuangyuan Molecular Biotechnology Corp.,Ltd was verified from four aspects:coincidence rate,precision,detection limit and anti-interference ability.Results Twenty clinical samples genotyping were detected both by quantitative PCR and Sanger sequencing simultaneously,the coincidence rate was 100%.The results of 15 times repeated detection in one clinical sample by quantitative PCR were in full accord,the Ct value of both CYP2C9 and VKORC1 gene fluorescence channels coefficient of variation(CV)were all<5%.The lowest detection limit of the kit was 10 ng/μL.After adding three interfering substances of hemoglobin,bilirubin and cholesterol into the blood samples with known genotype,and the results of the interference group and the control group were exactly the same.Conclusion The human CYP2C9 and VKORC1 gene polymorphism detection kit(quantitative PCR)meets the requirements of the relevant quality management of ISO15189 and can provide reliable test results for the clinic on the basis of ensuring project acceptance.
作者
李婧
李林静
刘转
杨文娟
马青梅
张安安
杨海棠
刘欣跃
LI Jing;LI Linjing;LIU Zhuan;YANG Wenjuan;MA Qingmei;ZHANG An′an;YANG Haitang;LIU Xinyue(Clinical Laboratory Center,Lanzhou University Second Hospital,Lanzhou,Gansu 730030,China)
出处
《检验医学与临床》
CAS
2021年第2期156-160,共5页
Laboratory Medicine and Clinic
基金
甘肃省青年科技基金计划项目(20JR5RA312)
兰州大学第二医院“萃英科技创新”计划项目(CY2018-QN06)
甘肃省教育厅创新能力提升项目(2019B-018)。
