免疫组织化学切片行EBER原位杂交检测方法的建立及应用  

Establishment and application of EBER in situ hybridization method on immunohistochemical section

作　　者：滕孝静[1] 程鸣[1] 周贺 王照情[1] 陈光勇[1] TENG Xiao-jing;CHENG Ming;ZHOU He(Department of Pathology,Beijing Friendship Hospital,Capital Medical University,Beijing 100050,China)

机构地区：[1]首都医科大学附属北京友谊医院病理科,北京100050

出　　处：《临床和实验医学杂志》2023年第15期1660-1664,共5页Journal of Clinical and Experimental Medicine

基　　金：国家重点研发计划项目(编号:2020YFC2004800)。

摘　　要：目的在石蜡组织免疫组织化学(IHC)阴性切片上建立EBER原位杂交(ISH)检测方法,为EB病毒相关淋巴组织病变在组织资源不足的情况下提供及时有效的检测结果及诊断依据。方法回顾性收集首都医科大学附属北京友谊医院病理科2021年1月至2023年2月间EBER原位杂交检测阳性的病例23例,其中NK/T细胞淋巴瘤8例,弥漫大B细胞淋巴瘤6例,伯基特淋巴瘤2例,经典霍奇金淋巴瘤4例,传染性单核细胞增多症3例。先选取1例组织量充足的EB病毒阳性弥漫大B细胞淋巴瘤组织用于IHC后EBER ISH前组织预处理条件的摸索,然后使用最佳预处理条件建立本实验的检测流程并用于另外22例组织中。22个蜡块每例连续切片2张,1张进行常规的EBER原位杂交检测,归为对照组;另1张归为实验组,先进行IHC染色,再按最佳预处理条件进行EBER原位杂交。最后对两组切片杂交的成功率、阳性强度、阳性细胞比例及组织细胞结构进行观察对比。结果实验组EBER杂交的成功率为90.91%(20/22),与对照组[100%(22/22)]相比,差异无统计学意义(P>0.05),杂交成功的切片阳性着色位于细胞核,定位准确,背景干净。两组切片的阳性细胞比例差异无统计学意义(P>0.05)。染色强度上,对照组中8例评分为3分的组织在实验组评分为2分,使两组染色强度的对比差异有统计学意义(P<0.05),但杂交结果仍可清晰明确地判读。实验组与对照组切片均未出现组织脱片、细胞形态结构不完整的现象。结论本文建立的IHC-EBER原位杂交检测方法操作简单、定位准确、成功率高、结果准确,可有效解决患者因切片或组织资源不足而无法进行EBER原位杂交检测的难题,能够为EB病毒相关淋巴组织疾病提供及时有效的诊断依据,具有临床实用价值,值得推广。Objective To establish the EBER in situ hybridization(ISH)method on the immunohistochemical(IHC)negative sections of paraffin tissue to provide timely and effective detection results and diagnosis basis for EB virus associated lymphoid tissue lesions in the case of insufficient tissue resources.Methods A total of 23 EBER-positive cases in the Department of Pathology,Beijing Friendship Hospital,Capital Medical University from January 2021 to February 2023 were retrospectively collected,including 8 cases of NK/T cell lymphoma,6 cases of diffuse large B cell lymphoma,2 cases of Burkitt lymphoma,4 cases of classical Hodgkin lymphoma,and 3 cases of infectious mononucleosis.One case of EBV-positive diffuse large B-cell lymphoma tissue with sufficient tissue volume was first selected to explore the pretreatment conditions of EBER ISH after IHC,and then the optimal pretreatment conditions were used to establish the detection process of this experiment and applied to the other 22 tissues.Two consecutive sections of each paraffin block were made.One was classified as control group.The other was classified as the experimental group to perform EBER in situ hybridization after IHC.Finally,the success rate,positive intensity,positive cell proportion and tissue structure of the two groups were observed and compared.Results The success rate of EBER hybridization in the experimental group was 90.91%(20/22),compared with the control group[100%(22/22)],and there was no statistically significant difference(P>0.05).There was no statistically significant difference in the proportion of positive cells between the two groups(P>0.05).In terms of staining intensity,8 tissues scored 3 points in the control group were scored 2 points in the experimental group,which made a statistically significant difference in the comparison of staining intensity between the two groups(P<0.05),but the hybridization results of 8 tissues could still be clearly interpreted.In both the experimental group and the control group,there were no dissections and inco

关 键 词：原位杂交 EB病毒 RNA 免疫组织化学 淋巴瘤 

分 类 号：R36[医药卫生—病理学]