Objective To prepare an internal quality control substance of hepatitis C virus RNA(HCV-RNA) used in real time quantitative PCR,and prepare quality control chart with Microsoft Office Excel software.Methods HCV-RNA positive serums were prepared,subpackaged into several centrifuge tubes and saved at -70 ℃.For the first time,all samples were continuously detected for 20 times in two tranches(group A:the first batch was detected under optimal conditions; group B;the second batch was detected under routine conditions).The logs of mean value(-x) ,standard deviation(s) and coefficient of variability(CV) were calculated and quality control chart was plotted.For the second time,the serums were stored at -70 ℃ for 12 months(group C;the third batch was detected under routine conditions).The logs of (-x), s and CV were calculated.Results The logs of (-x) of HCV -RNA internal quality control were 5.023 and 5.041,5 were 0.228 and 0.231,and CV were 4.54% and 4.58%.The difference in the logs of (-x) had no significance(P>0.05).There was also no statistical difference of s and CV.Conclusion Internal quality control substance of HCV-RNA used in real-time quantitative PCR might be very stable and applicable in clinical laboratory, and quality control chart could be plotted easily.%目的 自制HCV-RNA荧光定量PCR室内质控物,利用Excel绘制质控图.方法 取某一浓度HCV-RNA 阳性血清,稀释至一定浓度,分装数管,-70 ℃保存.首次分两批(A组:第1批在最佳条件下检测;B组:第2批在常规条件下检测)连续检测20次,计算均值的对数值、标准差和变异系数,绘制常规条件下质控图,进行室内质控动态监测;第2次(C组:第3批在常规条件下检测)标本-70 ℃保存12月后,连续检测20次,计算均值的对数值、标准差和变异系数.结果 HCV-RNA室内质控常规条件下及标本-70 ℃保存12个月后,均值的对数值分别为5.023、5.041;标准差分别为0.228、0.231;变异系数分别为4.54%、4.58%;经两组均数对数的显著性t检验,均值的对数值之间无统计学差异(P>0.05);标准差、变异系数之间相差很小.结论 荧光定量PCR 检测HCV-RNA质控物的制备方法简单,性能稳定,质控图绘制方便,适合PCR实验室对HCV-RNA荧光定量的室内质控.
展开▼
