全自动免疫组化与手工染色标记胸腺瘤TdT表达的对比观察

Comparative observation between automatic immunohistochemical staining technique and manual staining method in the TdT expression of thymoma

目的:对比观察胸腺瘤Td T在全自动免疫组织化学染色技术(automatic immunohistochemical staining techniques,AIST)和手工染色方法(manual staining method,MSM)中表达的差异性。方法:收集2015年-2016年大坪医院胸腺瘤手术标本21例,经石蜡包埋、切片、染色,显微镜下观察胸腺瘤Td T在AIST和MSM表达的差异。AIST采用Ventana公司生产的BenchM ark XT,二抗检测系统分别采用UltraV iew Universal DAB Detection kit(简称UV)和Optiview DAB IHC Detection kit(简称OV)。结果:手工染色TdT阳性表达定位准确,染色强度强,背景清晰;AIST中采用UV二抗检测系统TdT染色虽然阳性定位准确,但TdT染色强度偏弱,背景不清晰,两者比较具有显著性差异(P<0.01)。优化染色方案后,改用OV二抗检测系统,TdT染色各项指标优于UV二抗检测系统,染色效果和手工相比,染色强度和阳性率一致。结论:任何抗体指标在行免疫组化前均需进行最适条件的摸索与优化,从而获得该抗体指标最佳的染色方案。

Objective： The automatic immunohistochemical staining technique（ AIST） and manual staining method（ MSM） were used for comparison in the Td T expression of thymoma. Methods： All 21 cases of surgical thymoma FFPE specimens obtained from Daping hospital during 2015- 2016 were used to investigate the differences of AIST and MSM methods on Td T immunohistochemistry in thymoma. The Ultra View Universal DAB Detection kit（ UV） and Optiview DAB IHC Detection kit（ OV） were used respectively as the secondary antibodies on the Bench Mark XT for the AIST method. Results： The Td T protein detected by MSM showed accurate localization,strong staining intensity and clear background. However,the staining intensity and background of AIST used UV were not as satisfied as MSM,the comparison between the two groups had statistically significant defferences（ P〈 0. 01）. Therefore,the OV was used as the secondary antibody replacing the UV and showed equivalent results as MSM,with the staining intensity and background superior to UV. Conclusion： Novel antibodies（ both primary and secondary） should be optimized for the most appropriate protocol before used as the routine methods in the clinical immunohistochemistry practices.