两种前处理流程检测HBV DNA的重复性比较

Comparison of the coefficient of variation of HBV DNA detection by two kinds of preparation procedures

目的比较修改前后两种前处理流程检测HBV-DNA的变异系数(CV),以确定重复性更好的检测流程。方法采用修改前后两种前处理流程,对标称1×105IU/ml的阳性定量参考品、自制室内质控血清进行日内重复检测;对两个批号的自制室内质控血清进行日间重复检测,比较两种前处理流程检测结果的变异系数。结果以HBV DNA IU/ml的自然对数值进行统计分析,前处理流程修改前后,标称1×105的阳性定量参考品的日内CV分别为4.26%、1.38%,差异有统计学意义(u=2.564,P=0.0103);室内质控血清的日内CV分别为7.85%、2.14%,差异有统计学意义(u=2.792,P=0.0052);室内质控血清的日间CV分别为9.62%、3.40%,差异有统计学意义(u=7.358,P=1.9×10-13)。修改前处理操作流程后两个批号室内质控血清日间CV为3.40%、2.84%,差异无统计学意义(u=1.522,P=0.1281)。结论采用修改后的前处理流程检测HBV-DNA重复性更好。

Objective To compare the coefficients of variation（CVs）before and after the modification of the preparation procedure so as to determine the operation procedure to obtain better reproducibility.Methods Real-time PCR was used for the detection of HBV DNA.The preparation procedures before and after modification were used to repeatedly detect the positive reference quantified as ＂1×105＂ and the internal quality control serum in one day;the procedures were also used to repeatedly detect the internal quality control serum of two batches in different days.The CVs of the results were compared.Results Based on the log10 IU of HBV DNA/ml measured,before and after modification of the preparation procedure,the within-day CVs of the positive reference quantified as ＂1×105＂ were 4.26% and 1.38% respectively and the difference was significant（u=2.564,P=0.0103）;the within-day CVs of the internal quality control serum were 7.85% and 2.14% respectively and the difference was significant（u=2.792,P=0.0052）;the day-to-day CVs of the internal quality control serum were respectively 9.62% and 3.40% and the difference was significant（u=7.358,P=1.9×10-13）.The day-to-day CVs of the internal quality control serum of two batches detected after optimization were respectively 3.40% and 2.84% and the difference was not significant（u=1.522,P=0.1281）.Conclusion Use of the modified preparation procedure can obtain better reproducibility in the detection of HBV-DNA.