荧光定量PCR测定HBV DNA室内质控方法的探讨

目的探讨实时荧光定量聚合酶链反应(PCR)测定乙型肝炎病毒核酸(HBV DNA)的室内质控方法。方法统计上海地区PCR实验室HBV DNA常规条件下前20次室内质控数据的不精密度(CV),以测出的均值(x珋±s)绘制质控图,分别采用13s/22s的多规则质控方法和Levey-Jennings单规则质控方法,判断前20次室内质控数据CV分别为≥10%、5%~10%和1%~5%范围内A、B和C 3家实验室的室内质控数据,分别分析其失控检出能力。结果分别采用13s/22s多规则和Levey-Jennings单规则质控方法。当HBV DNA室内质控品低、高两个浓度分别为5×104和5×106IU/mL时,CV为14.96%和12.15%的A实验室均未正确检出失控数据;CV为6.49%和5.00%的B实验室检出2个随机误差引起的失控数据;其CV为4.36%和2.43的C实验室,采用13s/22s多规则质控方法检出3个系统误差引起的失控数据,采用单规则质控方法未检出失控。结论 PCR检测HBV DNA当实验室前20次室内质控数据CV≥10%时,无论采用单规则还是多规则质控方法均不能正确检出失控;实验室应设定自己实验室最低要求的CV,并采用多规则质控方法,以提高系统误差的失控检出率。

The investigation on the internal quality control of HBV DNA determination by fluorescence quantitation PCR

Objective To investigate the internal quality control of hepatitis B virus （HBV）DNA determination by real time fluorescence quantitation polymerase chain reaction （PCR）.Methods The coefficients of variation （CV）of previous 20 internal quality control data of HBV DNA determination under routine condition from PCR laboratories in Shanghai region were analyzed,and the control chart was drawn by the mean（-x±s）.1 3s/22s multi-rule quality control method and Levey-Jennings single-rule quality control method were used to determine internal quality control data from laboratory A,B and C whose CV were ≥1 0%,5%-〈1 0% and 1%-〈5%,respectively.Results When the concentrations of the internal quality control materials were 5 ×1 04 and 5 ×1 06 IU/mL,there were no error data detected in laboratory A whose CV were 1 4.96% and 1 2.1 5% by 1 3s/22s multi-rule quality control method and Levey-Jennings single-rule quality control method,respectively.There were both 2 random error data detected in laboratory B whose CV were 6.49% and 5.00% by 1 3s/22s multi-rule quality control method and Levey-Jennings single-rule quality control method,respectively.The CV of laboratory C were 4.36%and 2.43%.There were 3 systematic error data detected by 1 3s/22s multi-rule quality control method,and there were no error data detected by Levey-Jennings single-rule quality control method.Conclusions When the CV of the previous 20 internal quality control data were ≥ 1 0%,the error detections are all low both by the multi-rule quality control method and single-rule quality control method.The laboratory should set the suitable CV and use the multi-rule quality control method to improve the systematic error detection.