PML/RARα融合基因FISH检测试剂盒的研究  

Development of FISH diagnostic kit for detection of PML/RARα fusion gene in leukemia

作　　者：缪为民 石琳 胡林萍 李娟 刘芸 李承文 李思奇 杜宏伟 王征宇 MIAO Wei-min;SHI Lin;HU Lin-ping;LI Juan;LIU Yun;LI Cheng-wen;LI Si-qi;DU Hong-wei;WANG Zheng-yu

机构地区：[1]协和干细胞基因工程有限公司,天津300084 [2]中国医学科学院血液病医院/血液学研究所,天津300020 [3]天津市血液细胞治疗技术企业重点实验室,300084

出　　处：《中国医药生物技术》2018年第3期219-225,共7页Chinese Medicinal Biotechnology

基　　金：国家自然科学基金(81400150;81370598);天津市科委项目(14ZCDZSY00176);中国医学科学院医学与健康科技创新工程项目(2016-12M-1-017)

摘　　要：目的自主研制用于检测白血病染色体PML/RARα融合基因的双色荧光原位杂交(FISH)检测试剂盒。方法根据染色体基因图谱,选定合适的细菌人工染色体(BAC)克隆重叠群分别作为PML基因探针和RARα基因探针。对入选的BAC克隆进行STS-PCR鉴定和荧光原位杂交鉴定。最后将完成鉴定的PML探针标记红色荧光素,RARα探针标记绿色荧光素,加上配套试剂组成FISH检测试剂盒。用双盲法检测白血病临床样本37例,对自制探针和同类进口探针在各项技术参数上进行对比研究。结果自制试剂盒检测PML/RARα融合基因与进口探针相比,其阴、阳性率和同类进口产品完全符合;自制探针的荧光信号更强,信噪比更高。结论自主研制的PML/RARα融合基因FISH检测试剂盒设计合理、质量可靠,可用于替代价格昂贵的进口产品。Objective To develop a dual-colour FISH kit for detecting the leukemia PML/RARα fusion gene. Methods Based on the chromosome and gene map, corresponding BAC clone contigs were selected as PML and RARα FISH probes located on chromosome 15 q22 and 17 q21 respectively. The PML FISH and RARα probes were labeled with red and green fluorophores, respectively. The self-developed FISH kit and the commercial imported one were compared in terms of multiple technical parameters detected by double blind studies of 37 cases of clinical hematological samples. Results The detection results were consistent between the self-developed FISH kit and the commercial imported one with respect to detecting PML/RARα gene translocation. The self-developed probes were comparable to the commercial imported FISH probes in terms of multiple technical parameters, such as appearance, physical characteristics, fluorencence quench and stability at coldstorage. Furthermore, the self-developed probes possessed stronger fluorescence signal intensities and better signal noise ratios than the imported probes did. Conclusion The FISH diagnostic kit developed in this work could serve as a domestic replacement for the expensive imported products.

关 键 词：白血病 易位 遗传 原位杂交 荧光 PML/RARΑ融合基因 

分 类 号：R733.7[医药卫生—肿瘤]