荧光定量PCR测定HBVDNA的不确定度评定与应用  

Estimation and application of uncertainty of measurement in detecting of hepatitis B virus DNA by method of fluorescence quantitative polymerase chain reaction

作　　者：沈伟锋[1] 丁韧烨[1] 杨清萍[1] 邵平扬[1] 

机构地区：[1]浙江省嘉兴学院附属第一医院检验科,314000

出　　处：《中华检验医学杂志》2007年第2期169-172,共4页Chinese Journal of Laboratory Medicine

摘　　要：目的对荧光定量 PCR(FQ-PCR)测定 HBV DNA 测量不确定度进行评定,并探讨其应用价值。方法通过对 FQ-PCR 检测 HBV DNA 测量过程的分析,确定并简化测量不确定度的来源;采用不确定度 A 类评定方法以及不确定度 B 类评定方法,量化各不确定度分量;确定合成不确定度与扩展不确定度。结果测量不确定度来源主要有:批内重复性、批间重复性和方法偏倚等因素。FQ-PCR 测定 HBV DNA 的扩展不确定度 U=0.62(k=1.96,n=2);在各不确定度分量中,由方法偏倚导致的不确定度所占比重最大。结论 FQ-PCR 测定 HBV DNA 引入扩展不确定度使结果具有可比性,同时可作为乙型肝炎患者抗病毒治疗疗效判断和质控物浓度选择的依据。Objective To estimate the uncertainty of measurement in detecting of hepatitis B virus DNA（ HBV DNA） by method of fluorescence quantitative polymerase chain reaction （FQ-PCR） ,and discuss the application value. Methods The process of the detection of HBV DNA by FQ-PCR was analysed to confirm and simplify the sources of uncertainties of measurement, which were obtained by disposing the data of methodology validation, internal quality control （ type A evaluation of uncertainty） and external quality assessment（ type B evaluation of uncertainty） ; combined uncertainty and expanded uncertainty were obtained by statistical methods. Results The main sources of uncertainties of measurement were： precision within laboratory, precision between laboratory, method bias. The expanded uncertainty of HBV DNA by FQ-PCR was U=0. 62 （k =1.96,n =2）. The uncertainty caused by method bias was found mostly. Conclusion Expanded uncertainty can be compared in different results of HBV DNA by FQ-PCR , and it provides guide significance for observing the cure effect of anti-HBV and choosing the concentration of quality control.

关 键 词：不确定度 聚合酶链反应 肝炎病毒 乙型 DNA 

分 类 号：R450[医药卫生—治疗学]