实时荧光定量PCR快速检测BCR-ABL融合基因激酶区M244V突变方法的建立  

Establishment of Real-time Fluorescent Quantitative Polymerase Chain Reaction for Rapid Detection of M244V Mutation in Kinase Domain of BCR-ABL Fusion Gene

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作  者:黄颀[1,2,3] 杜新[1,2,3] 张巧霞[4] 卓家才[1,2,3] 

机构地区:[1]广州医科大学 [2]深圳大学第一附属医院 [3]深圳市第二人民医院血液科,广东深圳518035 [4]深圳大学第一附属医院,深圳市血液病研究所,广东深圳518035

出  处:《中国实验血液学杂志》2014年第6期1728-1734,共7页Journal of Experimental Hematology

基  金:广东省自然科学基金(S2012010008200)

摘  要:本研究建立一种敏感性好、特异性高的BCR-ABL融合基因M244V突变的检测方法。设计突变点特异性引物,优化实时荧光定量PCR反应体系和条件,建立检测BCR-ABL融合基因M244V点突变的定量PCR方法。结果表明,成功构建了M244V突变及野生的重组质粒标准品和M244V点突变的实时荧光定量PCR方法;验证结果表明该法具有较好敏感性、特异性和准确性。结论:BCR-ABL融合基因M244V突变的实时荧光定量PCR检测方法可用于临床CML患者M244V突变的检查。The present study was aimed to establish a high sensitive and specific method for detecting M244 V mutation in kinase domain of BCR-ABL( fusion gene)by using real-time quantitative PCR technology.The specific primer of the mutational site was designedand then the PCR reaction system and condition were optimized to establish the new real-time PCR method for detecting M244 V mutation.The results showed that a method of detecting M244 V mutation has been successfully established.The detection results indicated that this method possessed high sensitivityspecificity and accuracy.It is concluded that the method based on fluorescent quantitative polymerase chain reaction for detecting M244 V mutation can be used to detect the M244 V mutation in CML patients successfully.

关 键 词:BCR-ABL融合基因 M244V突变 实时荧光定量PCR 

分 类 号:R446.1[医药卫生—诊断学] R733.72[医药卫生—临床医学]

 

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