Wipe Test在HLA组织配型实验室污染监测及预防管理中的应用

目的建立人类白细胞抗原(HLA)组织配型实验室常规DNA污染监测试验体系,预防和消除可能会干扰检测准确性的PCR产物或基因组DNA的污染。方法设计含多种阴阳性对照和各关键监测点的完整反应链,针对HLA-Ⅰ类和Ⅱ类基因扩增区域内的非多态区进行引物特异性PCR,琼脂糖凝胶电泳测定扩增产物;对被污染监测点进行局部灭活处理后对相应监测点重新采样进行灭活验证;同时采取不定期抽查监测与定期全面监测相结合的方式确保实验过程零污染。结果灭活处理及整改后,所监测HLA基因分型实验过程中的核酸抽提区、PCR加样区、PCR扩增区、下游产物处理区4个区域的主要监测点污染监测结果均合格。结论该研究建立的以Wipe Test为核心的DNA防污染监测实验体系能够有效监测和灭活验证可能会干扰检测准确性的PCR产物或基因组DNA的污染,是HLA组织配型实验室质量控制管理程序的重要组成部分。

The application of Wipe Test in pollution monitoring and prevention management of HLA-typing laboratory

Objective To establish an experimental system of routine pollution monitoring of DNA in HLA tissue matching laboratories to prevent and eliminate contamination of PCR products or genomic DNA that might interfere with the accuracy of detection.Methods Design of a complete reaction chain containing multiple negative and positive controls and key monitoring points,and conduct prime-specific polymerase chain reaction for non-polymorphism regions in HLA-Ⅰand Ⅱ gene amplification regions,agarose gel electrophoresis for the determination of amplification products.Local inactivation was carried out on the contaminated monitoring point and the corresponding resampling was verified.At the same time,The combination of random spot check monitoring and regular comprehensive monitoring was adopted to ensure zero pollution in the experimental process.Results After inacation and rectification,the process of HLA gene typing experiments,the monitoring results of contamination of four main monitoring points in four regions,namely the extraction zone,the PCR sampling zone,the PCR amplification zone and the downstream product treatment zone were all qualified.Conclusion The DNA anti-pollution monitoring experimental system with Wipe Test as the core of this study can effectively monitor and inactivate the contamination of PCR products or genomic DNA that may interfere with the accuracy of the detection.It is an important part of the quality control management program of HLA organization matching laboratory.