儿童氨基糖苷类药物性耳聋基因快速无创检测法的应用

The application of a fast and noninvasive method for testing the mutations associated with aminoglycoside-induced hearing loss in children

目的:探讨氨基糖苷类药物性耳聋相关基因位点A1555G和C1494T的快速无创检测法在儿童耳聋诊断中的应用。方法:选取2015年5月至2017年5月温州医科大学附属第一医院诊断为非综合征型感音神经性耳聋患儿126例,收集其口腔黏膜拭子及静脉血。采用多重等位基因特异性PCR(MAS-PCR)检测口腔拭子来源DNA中A1555G和C1494T突变情况。同时用Sanger测序法检测静脉血来源DNA中A1555G和C1494T突变情况,并通过Kappa一致性检验分析2种方法检出率的一致性来验证前者的可靠性。结果:采用MAS-PCR检测口腔黏膜拭子来源DNA中A1555G和C1494T突变的检出率为6.35%(8/126),其中A1555G突变7例、C1494T突变1例。静脉血Sanger测序法的检出率为6.35%(8/126),Kappa一致性检验结果显示2种方法检出率一致性较好(kappa=1,P<0.01)。结论:采用MAS-PCR检测口腔黏膜拭子来源DNA中A1555G和C1494T突变是一种简便、快速、无创的检测手段,为婴幼儿,乃至新生儿药物性耳聋检测的开展提供一种可行的方法。

Objective： To probe the application of a fast and noninvasive method for testing the muta-tions A1555G and C1494T associated with aminoglycoside-induced hearing loss in children. Methods： A total of 126 non-syndromic sensorineural deafness patients aged 2 to 15 were selected for collecting oral swabs and venous blood from the First Affliated Hospital of Wenzhou Medical University during May 2015 to May 2017. Multiplex allele-specifc PCR was used to test the mutations of A1555G and C1494T in DNA from oral swabs. Simultaneously, Sanger sequencing was used to detect the mutations of A1555G and C1494T in the DNA of vein blood to verify the reliability of the former by Kappa testing. Results： The detection rate of A1555G and C1494T mutations in DNA from oral swabs detected by multiplex allele-specifc PCR was 6.35% （8/126）, including 7 cases of A1555G mutations and 1 cases of C1494T mutation. The testing rate of Sanger sequencing was also 6.35% （8/126）, and the two methods were in a good consistency checking （kappa=1, P〈0.01）. Conclusion： The testing of A1555G and C1494T mutations with multiplex allele-specifc PCR in DNA from oral buccal swabs is a simple, rapid and non-invasive method to provide another option for the testing of aminoglycoside-induced deaf-ness in infants and newborns.