广东省HER-2基因检测室间质控存在的问题分析

Analysis of the problems existed in the quality control of HER-2 gene testing in Guangdong Province

目的依托广东省临床病理质量控制中心分子病理组对广东省内医院开展的荧光原位杂交(fluorescence in situ hybridization,FISH)方法检测HER-2基因进行室间质控,全面了解HER-2基因检测的标准操作程序(standard operating procedure,SOP)的关键节点,以更好地指导乳腺癌靶向治疗。方法选择HER-2不同阳性模式的石蜡切片作为质控片,同一表达模式病例采取连续切片,首尾2张各做FISH检测以确保质控片质量的一致性。各参加实验室采用各自的荧光原位杂交法,根据实际情况选择相应的探针和消化酶等。质控结果由质控小组成员采取双盲形式进行结果判读。结果从我省参与HER-2基因检测质控的26家实验室汇报的结果来看,各实验室使用的试剂均为国家食品药品监督管理总局批准的试剂,3种试剂探针本身对质控结果的影响无明显差异;个别实验室仍存在一些细节操作及判读的问题,其中标本(1)结果吻合率96.15%,一家单位误判为簇状扩增,标本(2)结果吻合率84.62%,标本(3)结果吻合率100%;消化液使用了胃蛋白酶和蛋白酶K,2种酶的消化效果无特别明显差异,对质控片进行镜下观察发现有2家单位4′,6-二脒基-2-苯基吲哚(4′,6-diamidino-2-phenylindole,DAPI)通道下轮廓显示不清,提示存在消化过度的问题。结论各实验室应严格按照各实验室操作流程,保证染色过程中的温度、缓冲液及清洗液的浓度和p H值、探针的配置等参数准确。只有通过不断健全室间质控系统,科学设计基础与临床实验流程步骤和评价标准,配合孰知各项实验内容的检测团队,定期进行监查活动,才能确保提高我省HER-2检测的准确性和一致性。

Objective Relying on the molecular pathology group of Guangdong province clinical quality control center to carry out the external quality control for HER-2 gene detection by fluorescence in situ hybridization （FISH） methods in Guangdong province hospitals, in order to understand the standard operating procedure （SOP） key node of HER-2 gene detection entirely and better guide the targeted therapy of breast can- cer. Methods The paraffin sections with different positive patterns of HER-2 were selected as the quality control tablets, and the cases of the same expression pattern were performed consecutively, and the first and last 2 pieces were tested to ensure the consistency of quality control tablets. Each participating laboratories adopted the FISH methods of their own work and selected the corresponding probes and digestive enzymes according to the actual situation. The quality control results were interpreted by quality control team members in a double- blind manner. Results From the results of 26 laboratories participating in the quality control of HER-2 gene detection, all reagents used in the laboratories were approved by China Food and Drug Administration, and had no significant effect of these 3 probes on the quality control results. There were still some details of operation and interpretation in some individual laboratories. As a unit mistaken for cluster amplification, the self-agree- ment results of specimen （~） was 96.15%, specimen （~） was 84.62%, and the consistency of specimen （~） was up to 100%. The specimens were digested with Pepsin and proteinase K, and there were no obvious difference be- tween the 2 kinds of enzyme digestion. Meanwhile, microscopic examinations showed that DAPI staining were unclear in 2 units, it suggested the existence of excessive digestion. Conclusion Each laboratory should strictly follow the operating procedures of each laboratory to ensure that the temperature, buffer and cleaning liquid concentration and pH value, and probe configuration are accurate in the dyeing process. Only by constantly improving the external quality control system, the scientific design basis and clinical experiment process steps and evaluation standards, cooperating with the inspection team who knows the content, and regularly monitoring activities, the accuracy and consistency of HER-2 detection can be ensured in our province.