禽流感病毒检测免疫组织化学染色改良方法的建立

Establishment of an Improved Immunohistochemical Staining Method to Detect Avian Influenza Virus

目的 本研究对如何消除禽流感病毒检测免疫组织化学染色的非特异性反应进行探讨,从而建立一种免疫组化染色的改良方法。方法 对抗原修复方法、一抗孵育条件、内源性生物素封闭条件等进行优化,并用SABC检测系统对改良法和常规法进行比较。结果 采用微波炉加热法修复抗原、一抗在37℃孵育1.5～2 h、20%的蛋清液室温封闭20 min的改良法比常规法的染色效果好,可有效消除背景干扰,阳性表达清晰,对比度高。结论 建立的改良法特异性强,可用于禽流感病毒的基础研究和临床检验。

Objective In order to establish an improved immunohistochemical staining method to detect avian influenza virus, how to eliminate the nonspecific reaction was explored in the report. Methods The antigen retrieval, incubating condition of primary antibody, and blocking reaction of endogenous biotin were optimized and the results of improved method and the unimproved one were compared by SABC detection system. Results Satisfactory staining effect was obtained by antigen retrieval with microwave oven, primary antibody being incubated at 37 ℃ for 1.5 - 2.0 hours, and endogenous biotin being blocked by 20 % egg white solution for 20 minutes. Compared with common method, the improved one was able to eliminate the background interference effectively, and clear positive expression could be observed. Conclusion The improved method described was specificity enough and could be applied in preclinical medicine research and clinical detection for avian influenza virus.