国产HCV核酸扩增荧光定量试剂盒的溯源与单位统一的研究  

The tracability and uniform unit of hepatitis C virus RNA quantification by domestic made real-time fluorescence quantitative PCR method

作　　者：饶慧瑛[1] 季颖[1] 朱凌[1] 王江华[1] 刘峰[1] 魏来[1] 

机构地区：[1]北京大学人民医院北京大学肝病研究所,100044

出　　处：《中华检验医学杂志》2010年第10期909-912,共4页Chinese Journal of Laboratory Medicine

基　　金：国家＂十五＂科技攻关计划资助项目（2001BA705B06、2004BA718B10）;国家＂十一五＂科技重大专项资助项目（2008ZX10002-013）;北京大学人民医院研究与发展基金资助项目（RDC2007-18）

摘　　要：目的确定中国SFDA批准上市销售的国产HCV RNA扩增荧光定量试剂的量值X（copies/ml,lg）与标准物质的参考值Y（IU/ml,lg）之间的换算公式.方法用人AB型血清将WHO第二代HCV RNA国际标准品（编号：96/798）配制成不同浓度的样本7份,7份标准品的浓度分别为100 000、50 000、25 000、10 000、5 000、2 500、1 000 IU/ml.采用国内3种HCV RNA荧光定量试剂的2个不同的批次检测这7份不同浓度范围的标准品,每个批次重复测定4次.检测方法采用实时荧光定量PCR.结果 HCV RNA荧光定量试剂盒的量值X与标准品参考值Y之间的关系,试剂A：Y=0.902 0 X＋0.284 9,R2=0.953 3,P＜0.01,n=56;试剂B：Y=0.875 7 X＋0.562 4,R2=0.956 5,P＜0.01,n=56;试剂C：Y=0.843 8 X＋0.560 5,R2=0.945 8,P＜0.01,n=56.结论 3种试剂的量值与标准品参考值之间的换算公式不同,提示3种试剂的量值应进行更严格的溯源.3种试剂的换算公式能将其量值初步标准化,使不同试剂的量值之间具有可比性,为HCV感染的临床诊断和治疗监测初步提供统一的HCV RNA载量检测标准.Objective To determine the conversion equation for X（copies/ml, lg） quantity values of domestic HCV RNA quantitative fluorescence amplification assays approved by SFDA and Y（ IU/ml, lg）reference values of standard substance. Methods The second generation WHO International Standard （NIBSC code：96/798） was mixed with human AB blood type serum to create 7 different dilutions which included 100 000, 50 000, 25 000, 10 000, 5 000, 2 500 and 1 000 IU/ml. Two different batches of each three domestic hepatitis C virus RNA real-time fluorescence quantitative PCR assays and 2 different batches of each assay were employed to detect the 7 different concentration samples with real-time PCR. Each test was performed 4 times repeatedly. Results The correlations between X（ copies/ml,lg） values of domestic HCV RNA assays and Y（IU/ml,lg） reference values of standard substance were as follow,Assay A：Y =0. 902 0 X＋0.284 9,R2 =0.953 3,P〈0. 01,n =56;Assay B： Y=0. 875 7 X ＋0.562 4,R2 =0.956 5,P〈0.01,n =56; Assay C： Y = 0. 843 8 X ＋ 0. 560 5, R2 = 0. 945 8, P 〈 0. 01, n = 56. Conclusions All the conversion equations are different among the quantity value of three assays and the reference values of standard substance, that suggests it is necessary to perform more stringent traceability analysis for the quantity values of 3 assays. Through standardizing the quantity values preliminarily, the conversion equation can enhance the comparability between the quantity values of different assays, and provide a standard of HCV RNA virus load detection for clinical diagnosis and treatment monitoring of HCV infection.

关 键 词：肝炎病毒属 RNA 病毒 试剂盒 诊断 聚合酶链反应 

分 类 号：R686[医药卫生—骨科学]