荧光定量PCR测定HBV DNA测量不确定度评定的探讨

目的探讨利用质控数据评定荧光定量聚合酶链反应(PCR)测定HBV DNA测量不确定度的可行性。方法用批内不精密度和批间不精密度数据评定A类不确定度;分别用参加卫生部临床检验中心和美国病理学家协会(CAP)的室间质评数据评定B类不确定度;最后评定合成不确定度和扩展不确定度。结果利用卫生部临床检验中心室间质评反馈结果评定的扩展不确定度(k=1.96,n=1)在检测值为104~105和106~107IU/mL时分别为0.521 6和0.529 8;利用CAP能力比对反馈结果评定的扩展不确定度(k=1.96,n=1)在检测值为104~105和106~107IU/mL时分别为0.973 1和0.977 5。结论目前用室间质评数据评定PCR测定HBV DNA的不确定度不能真实反映实验室的不确定度,使用国家标准物质评定不确定度更合适。

Investigation on the evaluation of the uncertainty of hepatitis B virus DNA determination by fluorescence quantitative PCR

Objective To investigate the uncertainty of hepatitis B virus （HBV ） DNA determination by fluorescence quantitative polymerase chain reaction （PCR ） according to quality control data, and evaluate the feasibility.Methods The uncertainty of Type A was evaluated by the data of within-run and between-run coefficients of variation.The uncertainty of Type B was evaluated according to the data of external quality assessments from the National Center for Clinical Laboratory and College of American Pathologists（CAP）.The combined uncertainties and expanded uncertainties were evaluated and analyzed.Results The expanded uncertainties in 10^4-10^5 IU/mL and 106-107 IU/mL concentrations were 0.521 6 and 0.529 8（k=1.96,n=1）,according to the data from the National Center for Clinical Laboratory.The expanded uncertainties in 10^4-10^5 IU/mL and 10^6-10^7 IU/mL concentrations were 0.973 1 and 0.977 5 （k=1.96,n=1）,according to the data from CAP.Conclusions It is not bilievable to evaluate the uncertainty of HBV DNA determination by fluorescence quantitative PCR,and it is suitable to evaluate the uncertainty using the results traced back to the national standard substance.