应用荧光原位杂交技术诊断成人Ph样急性淋巴细胞白血病  

Diagnosis of adult Philadelphia chromosome-like acute lymphoblastic leukemia by fluorescence in situ hybridization

作　　者：林岱楠 黎秋丽 何显君 李环[1] 廖立斌[1] 何涵[1] 周玲玲 李珍 刘晓力[1] 刘启发[1] 周红升[1] 曹睿[1] Lin Dainan;Li Qiuli;He Xianjun;Li Huan;Liao Libin;He Han;Zhou Lingling;Li Zhen;Liu Xiaoli;Liu Qifa;Zhou Hongsheng;Cao Rui(Department of Hematology,Nanfang Hospital,Southern Medical University,Guangzhou,510515,China;Department of Hematology,the People's Hospital of Guangxi Zhuang Autonomous Region,Nanning,530000,China;Department of Hematology,General Hospital of Southern Theatre Command of PLA,Guangzhou,510010,China)

机构地区：[1]南方医科大学南方医院血液科,广州510515 [2]广西壮族自治区人民医院血液科,南宁530000 [3]解放军南部战区总医院血液科,广州510010

出　　处：《中华血液学杂志》2020年第9期749-755,共7页Chinese Journal of Hematology

基　　金：广东省医学科学技术研究基金(A2017307)。

摘　　要：目的建立应用荧光原位杂交技术(FISH)筛查成人Ph样急性淋巴细胞白血病(ALL)的体系。方法根据Ph样ALL的遗传学特征,设计了针对ABL1、ABL2、JAK2、EPOR、CRLF2、CSF1R、PDGFRB、P2RY8等基因断裂重排的FISH探针;对BCR-ABL1、ETV6-RUNX1、MLL基因断裂重排和E2A断裂重排均阴性的B-ALL,采用FISH进行Ph样ALL筛查,并结合流式免疫表型、靶向二代测序突变检测和RNA测序进行Ph样ALL诊断分析。结果2016年1月至2019年4月,南方医院血液科收治189例成人B-ALL,经FISH和(或)PCR检测,BCR-ABL1、ETV6-RUNX1、MLL断裂重排或E2A断裂重排阳性者共83例;其余106例患者接受Ph样ALL FISH探针筛查,其中,12例(11.3%)检出典型的Ph样ALL特异基因断裂重排,2例检出基因缺失。经RNA测序进一步验证,FISH检测Ph样ALL基因断裂重排结果灵敏度为71.4%,特异度为95.8%。综合免疫表型、靶向二代测序突变检测和RNA测序,共诊断融合基因阳性Ph样ALL 14例(13.2%)。结论FISH技术检测Ph样ALL具有较高的特异性,结合免疫表型和测序技术可完善诊断体系。Objective To establish a screening system of adult Philadelphia chromosome-like acute lymphoblastic leukemia(Ph-like ALL)by fluorescence in situ hybridization(FISH).Method Based on the genetic characteristics of Ph-like ALL,FISH probes were designed for ABL1,ABL2,JAK2,EPOR,CRLF2,CSF1R,PDGFRB,and P2RY8 gene breakpoints,which were used to screen Ph-like ALL in B-ALL patients without BCR-ABL1,ETV6-RUNX1,MLL,and E2A gene arrangement.Furthermore,it was analyzed in combination with flow immunophenotype,next-generation sequencing for targeted gene mutations,and RNA sequencing(RNA-seq).Results A total of 189 adult B-ALL patients diagnosed in Nanfang Hospital from January 2016 to April 2019 were enrolled in this study.Using FISH and/or PCR,BCR-ABL1,ETV6-RUNX1,MLL,or E2A arrangement was detected in 83 of them,and Ph-like ALL was detected by FISH in the other 106,resulting in the presence of typical gene arrangements of Ph-like ALL in 12 patients(11.3%,12/106).Validated by RNA-seq,the sensitivity and specificity of FISH for Ph-like ALL were 71.4%and 95.8%,respectively.After further analysis with immunophenotype,targeted gene mutations,and RNA-seq,14(13.2%,14/106)were diagnosed with Ph-like ALL.Conclusion This data shows high specificity of FISH for identification of Ph-like ALL and combining immunophenotype and sequencing technology can improve the diagnostic system.

关 键 词：荧光原位杂交 白血病 淋巴细胞 急性 Ph样 

分 类 号：R733.71[医药卫生—肿瘤]