荧光定量PCR测定HBV DNA的抗干扰能力评价

目的评价总胆红素、三酰甘油、血红蛋白和总免疫球蛋白G(IgG)对荧光定量PCR测定HBV DNA的干扰。方法参照EP7-A2方案对HBV DNA测定进行干扰评价试验。采用配对差异试验验证厂家的干扰声明,对不符合声明的干扰物通过剂量效应试验分析干扰物浓度与干扰程度之间的关系。结果配对差异试验结果显示476.0μmol/L总胆红素、34.0 mmol/L三酰甘油、2 g/dL血红蛋白对HBV DNA测定无干扰;40 g/L总IgG对HBV DNA测定有负干扰。剂量效应试验结果显示总IgG对HBV DNA测定可产生非线性负干扰。结论总胆红素、三酰甘油和血红蛋白对HBV DNA测定的干扰评价均符合厂家干扰声明;总IgG不符合声明。临床实验室应注意总IgG对HBV DNA测定的干扰,避免分析误差。

Evaluation of anti-interference ability of fluorescence quantitative PCR in detection of HBV DNA

Objective To evaluate the interference of total bilirubin(T-Bil),triglyceride,hemoglobin and total immunoglobulin G(IgG)for the results of fluorescence quantitative PCR in the determination of HBV DNA.Methods The evaluation of interference for fluorescence quantitative PCR in HBV DNA detection was tested according to EP7-A2 experimental procedures.The paired-difference test was applied to verify the claims of manufacturer on interference.The dose-effect of the interference materials incongruent with the manufacturer claim was analyzed to estimate the relationship between its concentration and interference degree.Results The results of paired-difference tests showed that 476.0μmol/L of T-Bil,34.0 mmol/L of triglyceride and 2 g/dL of hemoglobin did not exhibit interference effect on HBV DNA detection.However,40 g/L of total IgG showed negative interference effect on HBV DNA detection and the test results of dose-effect showed that total IgG produced non-linear negative interference effect on HBV DNA detection.Conclusion The interference evaluation of T-Bil,triglyceride and hemoglobin on HBV DNA detection meet with the interference claim of manufacturer,but the interference evaluation for total IgG presented opposite results.Clinical laboratory should pay attention to the interference effect of total IgG on HBV DNA detection to avoid the errors in analysis for test results.