用于荧光定量检测流感病毒RNA质控品的制备及质控图的绘制

Preparation of internal quality control substance of real-time RT-PCR to quantitative influenza virus RNA and drawing quality control chart

目的 应用自制流感病毒RNA室内质控品,绘制Levey-Jennings质量控制图.方法 选取近期的流感病毒流行代表株,提取RNA后10倍系列稀释（10-1 ～ 10-8）,取不同稀释度的RNA做RT-PCR荧光定量检测,每个稀释度平行做三孔,根据检测的平均Ct值选取高Ct值和低Ct值对应稀释度的RNA,连续检测20次,计算均值（x）和标准差（s）,应用Excel软件绘制Levey-Jennings质控图.结果 用Excel绘制出三种亚型流感病毒的Levey-Jennings质控图,三种亚型RNA质控品均在警戒线内.结论 三种亚型RNA质控品稳定性较好,制备简单,质控图绘制方便,适用于PCR检测实验室对流感病毒的室内质控.

Objective To draw a Levey-Jennings quality control chart using influenza virus RNA as internal quality control substance.Method Recent popular representative strains of influenza virus were selected and extracted RNA.Make series 10 folds dilution and take different dilution to do quantitative RTPCR.Three parallel wells of each dilution were done.High Ct value and low Ct value corresponding dilution of RNA were selected and detected continuously twenty times.Mean Ct value （x） and standard deviation （s） were calculated.Levey-Jennings quality control chart were drew by using Excel software.Result Levey-Jennings quality control charts of three influenza virus subtype were drew and each internal quality control substance was in control.Conclusion RNA quality substance of each subtype has good stability and preparation simple which is convenient to draw quality control chart and suitable as internal quality control substance for PCR laboratory.