循环荧光原位杂交在淋巴瘤诊断中的价值研究  

The application of recyclable fluorescence in situ hybridization in the diagnosis of lymphoma

作　　者：滕孝静[1] 刘伟[1] 毕阔 孙岚[1] 王照情[1] 杨璐晶 TENG Xiao-jing;LIU Wei;BI Kuo(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》2022年第13期1345-1348,共4页Journal of Clinical and Experimental Medicine

基　　金：国家重点研发计划“老年人多病共患临床大数据与生物样本库综合管理共享平台建设”项目资助(编号:2020YFC2004800)。

摘　　要：目的在单张淋巴瘤石蜡切片上建立循环荧光原位杂交检测的方法,为需要多基因检测的淋巴瘤患者在组织或切片不足的情况下提供及时有效的诊断。方法回顾性收集2020年1月至2021年5月首都医科大学附属北京友谊医院病理科(北京市临床医学研究所淋巴瘤诊断研究中心)和外院会诊的淋巴瘤病例共27例,其中本院10例,外院会诊17例。27例均进行过MYC、BCL2和BCL63种基因的FISH检测,每例切片3张。根据杂交的不同将实验分为3组:杂交1组(HG1):将3张切片分别滴加MYC、BCL2、BCL6探针并按我科室常规荧光原位杂交方法操作。杂交2组(HG2):将HG1组中杂交MYC探针的切片使用本文介绍的循环荧光原位杂交方法,滴加BCL2探针进行第2次杂交。杂交3组(HG3):将HG2组的切片用循环方法滴加BCL6探针进行第3次杂交。观察3组切片红、绿荧光信号的有无、信号模式及组织、细胞结构情况。结果HG2组和HG3组循环荧光原位杂交的成功率均为100%。循环杂交后,HG2组中BCL2基因和HG3组中BCL6基因断裂阳性、阴性及扩增病例与HG1组完全相符,每张切片上基因表达异常的细胞数目与HG1组相比,差异无统计学意义(P>0.05)。HG2组和HG3组在循环杂交后,切片上的组织结构完整,细胞轮廓清晰,信号明亮清晰、定位准确。结论循环荧光原位杂交检测法可以在同一张切片上多次杂交不同的基因探针,方法简便,成功率高,结果准确,可有效解决淋巴瘤患者因组织或切片不足而无法进行多基因检测的问题,在淋巴瘤诊断中具有实用价值。Objective A method of recyclable fluorescence in situ hybridization detection was established on a single lymphoma paraffin section to provide timely and effective diagnosis for lymphoma patients who need multi-gene detection in the case of insufficient tissue or sections.Methods A total of 27 lymphoma cases were retrospectively collected at the Department of Pathology,Beijing Friendship Hospital,Capital Medical University(Lymphoma Diagnosis Research Center,Beijing Institute of Clinical Medicine)from January 2020 to May 2021.Among them,10 cases were in our hospital and 17 cases were consulted from other hospitals.All 27 cases underwent FISH detection of MYC,BCL2 and BCL6 genes,and each case had 3 sections.According to the difference of hybridization,the experiment was divided into 3 groups:Hybridization group 1(HG1):MYC probe,BCL2 probe and BCL6 probe were hybridized to the three sections respectively according to the routine fluorescence in situ hybridization method in our laboratory.Hybridization group 2(HG2):Sections in the HG1 group hybridized with the MYC probe were hybridized with BCL2 probe for the second hybridization using the recyclable method described herein.Hybridization group 3(HG3):Sections of the HG2 group were hybridized with BCL6 probe by the recyclable method for the third hybridization.The presence or absence of red and green fluorescence signals in the three groups of sections,the signal patterns and the structure of tissue and cells were observed.Results The success rate of recyclable fluorescence in situ hybridization was 100%in both HG2 and HG3 groups.After recyclable hybridization,the number of positive,negative and amplified BCL2 gene cases in HG2 group was consistent with that in HG1 group,and there was no significant difference in the number of cells with abnormal gene expression between HG2 group and HG1 group(P>0.05).The number of positive,negative and amplified BCL6 gene cases in HG3 group was consistent with that in HG1 group,and there was no significant difference in the number of

关 键 词：原位杂交 荧光 循环 淋巴瘤 

分 类 号：R733.1[医药卫生—肿瘤]