杂交前3%过氧化氢预处理增强EBER原位杂交特异性阳性信号

Pretreatment with 3%hydrogen peroxide before hybridization enhances specific positive signal of EBER in situ hybridization

目的优化EBER原位杂交检测方法,增强特异性阳性信号,降低背景染色。方法选取30例EBV阳性的组织切片,在杂交前处理时加入3%过氧化氢,再通过EBER探针进行原位杂交,比较不同过氧化氢孵育时间对杂交反应阳性信号和背景的影响。结果用3%过氧化氢处理鼻咽癌组织10 min、处理淋巴瘤组织15 min,EBER原位杂交阳性信号强,背景干净。结论EBER原位杂交过程中,在杂交前处理时用3%过氧化氢短暂孵育可以增强特异性阳性信号,降低非特异性背景染色。

Objective Optimize EBER in situ hybridization method to enhance specific positive signals and reduce background staining.Methods 30 EBV positive tissue sections were selected,and 3%hydrogen peroxide was added during pre-hybridization treatment,and EBER probe was used for in situ hybridization.The influence of different incubation time of hydrogen peroxide on the positive signal and background of hybridization reaction was compared.Results Treatment with 3%hydrogen peroxide of nasopharyngeal carcinoma tissue for 10 minutes and lymphoma tissue for 15 minutes showed strong positive signals of EBER in situ hybridization and a clean background.Conclusion During the EBER in situ hybridization process,short-term incubation with 3%hydrogen peroxide before hybridization can enhance specific positive signals and reduce non-specific background staining.