改良手工免疫组化方法和改良原位杂交方法的应用研究

Application of improved manual immunohistochemistry and improved in situ hybridization

目的探讨改良手工免疫组化染色和原位杂交方法的实用价值和推广意义。方法免疫组化:选取本院近期病例80例,每例分别进行常规手工和改良手工染色及全自动免疫组化仪染色,进行结果对比;原位杂交:选取本院近期原位杂交EBER阳性病例15例和HPV阳性病例18例,每例分别进行传统和改良手工原位杂交染色方法,并比较染色结果。改良手工染色方法为:省去一抗后的清洗步骤,直至显色前充分清洗。结果三种免疫组化染色方法均定位准确,无非特异性染色,待测目的组织与内对照的表达情况一致,复染清晰,染色评分无差异(P=0.985),但全自动免疫组化仪的染色背景更加干净。三种方法的脱片率分别为18.1%、17.8%和10.0%,传统手工和改良手工免疫组化方法之间脱片率差异无统计学意义(P=0.911),传统手工与全自动免疫组化仪染色方法之间、改良手工与全自动免疫组化仪染色方法之间脱片率差异有统计学意义(P=0.009,P=0.012)。传统手工方法和改良手工方法原位杂交染色的结果完全一致。结论免疫组化和原位杂交的改良手工染色法可以代替传统手工染色法。

Objective To explore the practical value and promotion significance of the improved manual immunohistochemical staining and the improved in situ hybridization method. Methods A total of 80 recent cases in Beijing Chao-Yang Hospital were selected,and each case was stained with traditional manual immunohistochemical staining,modified manual immunohistochemical staining and automatic immunohistochemical instrument;15 EBV in situ hybridization positive cases and 18 HPV in situ hybridization positive cases were selected,and each case was stained with traditional manual method and modified manual method,and the results were compared. Results The three methods of immunohistochemistry staining were all located accurately,there were no non-specific staining,the expression of the target tissue were consistent with internal control,the results of counterstaining were clear,and the difference of the staining scores was not statistically( P = 0. 985),but the staining backgrounds of the fully automated immunostaining group were clearer. The stripping rates of the three methods were 18. 1%,17. 8% and 10. 0%,respectively. There was no statistical difference between the traditional manual immunohistochemical staining and the modified manual immunohistochemical method( P = 0. 911),but the stripping rates between the traditional manual immunohistochemical staining and the automatic immunohistochemical staining and the stripping rates between the improved manual immunohistochemical staining and automatic immunohistochemistry staining were statistically significant( P = 0. 009,P = 0. 012). The results of in situ hybridization with traditional manual methods and improved manual methods were completely consistent. Conclusion The improved manual staining method of immunohistochemistry and in situ hybridization which simplify the operation steps and shorten the operation time can replace the traditional manual staining methods. They are low-cost and are worthy to promote in primary hospitals.