[1]付朝泓,李艳,童永清,等.BCR-ABL融合基因核酸检测试剂的研发及临床应用[J].现代检验医学杂志,2015,30(06):52-55.[doi:10.3969/j.issn.1671-7414.2015.06.015]
　FU Chao-hong,LI Yan,TONG Yong-qing,et al.Development and Clinical Application of A Novel Nucleic Acid Detection Reagent for BCR-ABL Fusion Gene[J].Journal of Modern Laboratory Medicine,2015,30(06):52-55.[doi:10.3969/j.issn.1671-7414.2015.06.015]
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BCR-ABL融合基因核酸检测试剂的研发及临床应用()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第30卷
期数:
2015年06期
页码:
52-55
栏目:
论著
出版日期:
2015-12-30

文章信息/Info

Title:
Development and Clinical Application of A Novel Nucleic Acid Detection Reagent for BCR-ABL Fusion Gene
作者:
付朝泓; 李艳; 童永清; 陈占国
武汉大学人民医院检验科，武汉430060
Author(s):
FU Chao-hong; LI Yan; TONG Yong-qing; CHEN Zhan-guo
Department of Clinical Laboratory，Renmin Hospital of Wuhan University，Wuhan 430060，China
关键词:
BCR-ABL; 融合基因; 荧光定量; 白血病
分类号:
Q789
DOI:
10.3969/j.issn.1671-7414.2015.06.015
文献标志码:
A
摘要:
目的研发同时检测多种BCR-ABL融合基因类型的试剂并建立相对应的荧光定量检测方法，同时进行临床标本检测。方法检索GeneBank等生物信息数据库，获得BCR-ABL基因可能的融合亚型，对亚型之间的序列进行比对并设计BCR-ABL融合基因特异性引物，从而应用质粒建立新的荧光定量体系。收集武汉大学人民医院2014年1月1日～11月31日住院部确诊为初发白血病患者30例，其中慢性髓细胞白血病15例，急性淋巴细胞白血病15例，另同时收集体检中心健康对照20例。应用新建立体系、商品试剂盒及直接测序法进行BCR-ABL融合基因的检测。结果新建荧光定量PCR体系准确度验证结果阳性符合率为100%，阴性符合率为100%；特异性验证结果各干扰项检测均为阴性；可检测11种融合基因类型。对BCR-ABL融合基因亚型e13a2，e14a2和e1a2的检测新方法阳性符合率为100%(10/10)，阴性符合率为100%(5/5)。对稀有融合基因亚型e19a2检测新方法检测阳性率100%(2/2)，商品试剂盒检测阳性率为0%(0/2)。新方法与直接测序法相比阳性符合率为100%(16/16)，阴性符合率为100%(14/14)。结论以研发的BCR-ABL融合基因检测试剂而建立的荧光定量方法检测BCR-ABL融合基因与测序结果符合率为100%，是一种快速、可靠的检测方法，可应用于临床。
Abstract:
ObjectiveTo developand measure the type of BCR-ABL fusion genes and established the detection method of the fluorescent quantitative polymerase chain reaction and detect the clinical samples by new method．MethodsThe subtype of BCR-ABL fusion gene would be obtain by bio information databases and compared differently among them．The specific primer of BCR-ABL fusion gene were designed and the new system of fluorescent quantitative testing were created using plasmid．Collected 30 leukemia cases of inpatient department of Renmin Hospital of Wuhan University from January to November 2014，including 15 cases of chronic myeloid leukemia and 15 cases of acute lymphoblastic leukemia．Used new method，commercialized kit and direct sequencing to test clinical samples．ResultsThe negative result and the positive result of new PCR system were all 100%．The detecting result of disturbance term was negative on side of specificity．The new method could screen 11 subtypes of BCR-ABL fusion gene．The negative result and the positive result of e13a2，e14a2 and e1a2 accorded with commercialized kit were 100%(10/10)，100%(5/5)，respectively．The positive result of e19a2 be different with commercialized kit were 100%(2/2)，0%(0/2)，respectively．The negative result and the positive result of subtypes of BCR-ABL fusion gene accorded with direct sequencing were 100 %(16/16)，100%(14/14)，respectively．ConclusionsThe testing result of new method was compared accuracy reaching 100% with the data of direct sequencing．This detecting method is quick，effective and reliable and would be applied clinical．

参考文献/References:

［1］谷红丽，王进，秦正红，等．慢性粒细胞白血病BCR-ABL融合类型的研究及其临床意义［J］．苏州大学学报(医学版)，2012，32(1)：84-88． Gu HL，Wang J，Qin ZH，et al．Types of BCR-ABL fusion gene in chronic myeloid leukemia and its clinical significance［J］．Suzhou University Journal of Medical Science，2012，32(1)：84-88．
[2]赵晶晶，王颖，张文伟，等．使用Taqman技术监测慢性粒细胞白血病患者微小残留病灶［J］．分子诊断与治疗杂志，2014，6(4)：239-244． Zhao JJ，Wang Y，Zhang WW，et al．Detection of minimal residual in chronic myelocytic leukemia (CML) patients by Taqman technique［J］．J Mol Diagn Ther，2014，6(4)：239-244．
[3]Burmeister T，Maurer J，Aivado M，et al．Quality assurance in RT-PCR-based BCR/ABL diagnostics-results of an interlaboratory test and a standardization approach［J］．Leukemia，2000，14(10)：1850-1856．
[4]Burmeister T，Reinhardt R．A multiplex PCR for improved detection of typical and atypical BCR-ABL fusion transcripts［J］．

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备注/Memo

备注/Memo:
作者简介：付朝泓(1978-)，男，硕士，主管技师，研究方向：分子诊断。 通讯作者：李艳，E-mail：yanlitf1120@163.com。
更新日期/Last Update: 1900-01-01