eber原位杂交结果的稳定性与变性温度的研究.docVIP

精品论文 参考文献 EBER原位杂交结果的稳定性与变性温度的研究 （皖南医学院弋矶山医院病理科 芜湖 241001） 【摘要】目的:探讨原位杂交变性温度的改变对EBER阳性表达的影响。方法：采用不同变性温度对多例EBER阳性表达的标本进行原位杂交检测。结果：A组50℃：整体几乎阴性，几张微弱阳性，B组55℃：阳性较A组多，强度不稳，整体不均一，C组60℃：基本阳性，强度较强，有几例强度偏弱，D组70℃：全部阳性，强度强，稳定，均一。E组75℃：全部阳性，强度强，背景增强，假阳性率高，特异性降低。结论：原位杂交检测EBER阳性表达在一定范围里随变性温度升高信号稳定性增强，70℃为最佳变性温度。 【关键词】 原位杂交；EBER；温度 【中图分类号】R3 【文献标识码】A 【文章编号】2095-1752（2015）35-0375-02 Study on the stability of EBER in situ hybridization and the temperature of the modified LI jiajia,XU Guoxiang, HE Lei, MEI Jingjing, ZHANG Wei-uan, CHENG Jiao(Department of pathology, Yijishan Hospital, Wannan Medical College, Wuhu,China, 241001) 【Abstract】Objective: To investigate the effect of the change of the temperature of the in situ hybridization on EBER expression. Method: In situ hybridization detection was performed by using different temperature to detect the positive expression of EBER. Result: A group of 50 ℃: almost the whole negative, a weakly positive, B group 55 ℃: compared with the positive group a multi, intensity is unstable, as a whole is not uniform, and group C 60 ℃: was basically and strong intensity, a few cases of strength weak, group D 70℃: all positive, strength, strong, stable, uniform. The E group was 75℃, all positive, strong intensity, background enhancement, high false positive rate, specificity decreased. Conclusion: In situ hybridization detection of EBER positive expression in a certain range, with the increase of the temperature of the modified signal stability enhancement, 70℃ for the optimum temperature of denaturation. 【Key words】In situ hybridization; EBER; temperature Epstein-Barr病毒(EBV)与鼻咽癌(nasopharyngeal carcinoma， NPC)非常密切的关系。各家检测EBV DNA的方法均不完全相同，灵敏度也存在差异。原位杂交是比较传统的方法，但用EBV-DNA 片段作探针原位杂交的检测灵敏度并不高，而且检测步骤相对繁琐。我们用EBER(Epstein-Barr virus encoded RNAs)-1作探针原位检测鼻咽癌组织中的EB病毒，获得了满意的效果。 1.材料和方法 1.1材料 1.1.1标本 鼻咽部活检组织石蜡包埋切片标本由安徽省皖南医学院附属弋矶山医院病理科提供，共25例，均为近1年内存档检测过阳性标本，按常规福尔马林固定、石蜡包埋、连续切片(2-4mu;m