解脲支原体DNA检测室内质控物的制备及质控方案的设计

Preparation of internal quality control substance of real-time PCR to analyze UU-DNA and the design quality control programs

目的制备解脲支原体（UU）DNA荧光定量PCR室内质控物,建立室内质量控制体系,对其临床应用进行初步评价。方法分别留取Ct值为24~25（阳性）和32~33（弱阳性）标本和检测结果为阴性的标本,待留取到15mL时充分混匀,按每管150μL分装作为室内质控物。前20次检测采用＂即刻法＂判断检测结果是否在质控范围内,20次以后绘制Levey-Jennings图,确定靶值、标准差（s）和变异系数（CV）,采用Westdard多规则质控方法对自制室内质控物检测结果进行判断。在Unity Real Time（URT）系统中使用质控规则配置操作导出UU-DNA的功效函数图（OPSPecs图）,根据OPSPecs图设置质控规则。结果131次实验中,前20次采用＂即刻法＂判断室内质控物;20次以后采用Levey-Jennings图判断,质控品稳定,质控规则合理。结论用临床分泌物混合液制备UU-DNA检测项目的室内质控物品,制备方法简单,测定结果稳定,可作为实验室检测UU-DNA项目的质控品;依据OPSPecs图设置分子生物学检测项目的室内质控规则简便、实用。

Objective Developing an internal quality control substance of Ureaplasma urealyticum（UU）-DNA for real-time PCR to establish an internal quality control system and preliminary evaluation its clinical value.Methods Internal quality control substance was prepared by mixing samples which Ct value were 24-25（positive sample）and 32-33（weak positive sample）,respectively.At the same time,selecting samples that test results were negative as negative control.The target value,standard deviation（s）and coefficient of variation（CV）of internal quality control substance were defined by＂instant method＂for the first 20 runs and Levey-Jennings quality control（QC）chart after the first 20 runs.Using the＂Westdard＂multi-rule quality control methods to analyze the detection results.Exporting OPSPecs chart by quality control rules in Unity Real Time（URT）system and setting up new quality control rules according with OPSPecs chart.Results 131 times of the detection of quality control substance were performed totally.The first 20 runs were defined by＂instant method＂and later 111 runs were defined by Levey-Jennings QC chart,the results were stable of quality control substance and reasonable quality control rules.Conclusion Preparing of internal quality control substance of UU-DNA used in real-time PCR might be easy and stable.So,the internal quality control substance of UU-DNA could be worthy for practical application in this PCR laboratory.Design internal quality control rules based OPSPecs chart in molecular detection is very simple and practical.