荧光原位杂交和原位杂交联合免疫荧光检测EBER的可行性比较

The comparison and feasibility analysis of fluorescence in situ hybridization and in situ hybridization combined with immunofluorescence for EBER detection

目的探讨利用荧光原位杂交(FISH)和原位杂交(ISH)联合免疫荧光(IF)两种方法检测EB病毒编码的小RNA(EBER)的可行性。方法收集首都医科大学附属北京友谊医院病理科2016年7月至2017年1月诊断的淋巴瘤患者20例,EBER原位杂交检测均为阳性。每例取蜡块进行切片,每张厚3μm,4张连续切片分为四组:荧光原位杂交及其阴性对照,原位杂交联合免疫荧光及其阴性对照。通过荧光显微镜查看两种荧光方法的效果。结果利用荧光原位杂交与原位杂交联合免疫荧光的方法检测EBER,均可以使EBER阳性细胞显示出阳性绿色荧光信号,且阴性对照无阳性荧光信号。后者更敏感,背景更少。结论荧光原位杂交与原位杂交联合免疫荧光这两种方法都可以有效地应用于EBER的检测,且这两种方法可能会成为EB病毒相关疾病诊断的实验室检查方法。

Objective In order to find the feasibility of using fluorescence in situ hybridization（ FISH） and in situ hybridization（ ISH）combined with immunofluorescence（ IF） for EBV-encoded RNA（ EBER） detection. Methods Twenty cases of patients with EBV-positive expression were tested in this study. These twenty cases were diagnosed with lymphoma from July 2016 to January 2017 in department of pathology in Beijing Friendship Hospital. The specimen of each case was sectioned into 3-micron fragments. 4 sequential sections were divided into four groups： FISH,negative control for FISH,ISH combined with IF,negative control for ISH combined with IF. The reading was done on fluorescence microscopy. Results Both FISH and ISH combined with IF could show positive green fluorescence on fluorescence microscopy while the negative control had no fluorescence but the background. And ISH combined with IF was more sensitive with less background. Conclusion We can consider FISH and ISH combined with IF to be sensitive and specific methods to detect EBER. They both can be auxiliary laboratory tools in the diagnosis of EBV infection.