TaqMan探针熔解曲线技术检测GJB2基因突变

Diagnosis of hereditary deafess with GJB2 gene mutations based on TaqMan probe melting curve technology

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。

Objective To establish a quick and reliable Taq Man probe melting curve technology to be used for analyzing the GJB2 gene in patients with non-syndromic hearing impairment（NSHI）. Methods Standard samples were prepared to establish the Taq Man probe melting curve technology based on q PCR. Samples were then collected from 138 normal individuals, 113 patients and 2 families with NSHI to test 35 del G,176_191del16, 235 del C, 299_300del AT mutations of the GJB2 gene using the Taq Man probe melting curve technology. Test results were verified by direct DNA sequencing. Result Two of the 138 normal individuals, 22 of the113 NSHI patients and both NSHI families were shown to carry GJB2 gene mutations. All results were consistent with those by direct DNA sequencing. Conclusion A quick and reliable Taq Man probe melting curve technology has been established and shown to be capable of detecting 4 mutations of the GJB2 gene.