TaqMan探针熔解曲线技术检测GJB2基因突变

发布时间：2018-11-10 07:51

【摘要】：目的建立快速可靠的Taq Man探针熔解曲线技术,分析非综合征型遗传性耳聋患者的GJB2基因及探讨Taq Man探针熔解曲线技术。方法制备标准品,利用Taq Man探针基于荧光PCR熔解曲线平台建立熔解曲线分析技术;收集138例正常人群、113例非综合征型遗传性耳聋患者及2个非综合征型遗传性耳聋家系,应用Taq Man探针熔解曲线技术分析GJB2基因最常见的35del G,176_191del16,235del C,299_300del AT的4个突变位点;结果利用直接测序技术加以验证。结果 138例正常人群中检出2例GJB2基因突变,113例非综合征型遗传性耳聋患者中检出22例GJB2基因突变,2个耳聋家系先证者均为GJB2基因突变患者;所有检测结果均与测序结果一致。结论利用Taq Man探针建立了一种快速可靠的探针熔解曲线技术,经测序验证能准确的检测GJB2基因常见的4种突变。
[Abstract]:Objective to establish a rapid and reliable Taq Man probe fusion curve technique to analyze the GJB2 gene in patients with non-syndromic hereditary deafness and to explore the Taq Man probe fusion curve technique. Methods the standard sample was prepared and the technology of melting curve analysis was established by using Taq Man probe based on fluorescence PCR melting curve platform. 138 normal subjects, 113 non-syndromic hereditary deafness patients and 2 families with non-syndromic hereditary deafness were collected and analyzed by Taq Man probe fusion curve technique. 4 mutation sites of 299_300del AT; Results the results were verified by direct sequencing technique. Results 2 cases of GJB2 gene mutation were detected in 138 normal subjects, 22 cases of GJB2 gene mutation were detected in 113 cases of non-syndromic hereditary deafness, and 2 cases of proband of deafness were GJB2 gene mutation. All the results were consistent with the sequencing results. Conclusion A fast and reliable method of probe melting curve was established by using Taq Man probe, and four common mutations of GJB2 gene could be detected accurately by sequencing.
【作者单位】：暨南大学第二临床医学院深圳市人民医院;亚能生物技术(深圳)有限公司;
【基金】：2013年深圳市战略新兴产业发展专项资金(CXZZ20130517143111780)
【分类号】：R764.43

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