实时荧光PCR的探针熔解曲线分析与质谱分析应用于耳聋基因诊断的对比研究  

Comparison between the probe melting curve analysis based on real-time fluorescence PCR and Mass Spectrometry for gene diagnosis of deafness

作　　者：王格[1] Wang Ge(Zhuhai Municipal Maternity and Child Healthcare Hospital,Zhuhai, Guangdong 519000, China)

机构地区：[1]珠海市妇幼保健院,广东珠海519000

出　　处：《医药前沿》2019年第27期28-30,共3页Journal of Frontiers of Medicine

摘　　要：目的:评价基于实时荧光PCR的探针熔解曲线分析(PMCA)技术作为耳聋基因基因诊断的可行性.方法:选取2017年1-4月在珠海市妇幼保健院出生的经MALDI-TOF-MS诊断为耳聋基因携带者的婴儿的脐带血标本92份和正常的脐带血标本4份,采用PMCA技术对上述96份标本进行耳聋基因检测,检测结果不一致的样本,以DNA测序技术作为金标准,对上述两种方法检测耳聋基因突变的检测性能进行评估.结果:探针熔解曲线分析法除了2例不在检测范围外,其余的94份标本均能正确检出.经测序鉴定,该两例未能检测出,是由于基因检测位点的不同所导致.结论:PMCA技术可作为MALDI-TOF-MS技术检测耳聋基因基因突变的替代或验证方法.Objective To evaluate the reliability of the probe melting curve analysis(PMCA)based on real-time fluorescence PCR for gene diagnosis of deafness.Methods From January to April 2017,92 umbilical cord blood samples and 4 normal Umbilical cord blood samples from infants diagnosed as deaf gene carriers by MALDI-TOF-MS were collected from Zhuhai Maternal and Child Healthcare Hospital.The PMCA technique was used to detect the deafness gene in 96 samples.The DNA sequencing technique was used as the gold standard to evaluate the detection performance of the above two Methods for detecting the gene mutation of deafness.Results The genotypes of 96 Cases were detected aceuralely based on the PMCA assay except for two cases:Two cases were not in the detection range,Sequencing confirmed that the two cases could not be detected because of the differences in gene detection sites.Conclusion PMCA technique can be used as a substitute or verification method for detecting gene mutation of deafness by MALDI-TOF-MS.

关 键 词：耳聋基因诊断 熔解曲线分析 质谱分析 

分 类 号：R764.43[医药卫生—耳鼻咽喉科]