探讨聚合酶链反应-反向点杂交(PCR-RDB)法对遗传性耳聋相关基因突变检测的准确性和实用性。

选择2014年1月至2016年10月，于广东省妇幼保健院临床诊断为非综合征型耳聋(NSHI)的250例患者为研究对象。采用PCR-RDB法检测其外周血遗传性耳聋相关基因突变。PCR-RDB法采用遗传性耳聋基因芯片检测试剂盒，可检测中国人群常见的4个耳聋相关基因的12个热点突变，即GJB2(35del G、176_191del 16、235del C、299_300del AT)，GJB3(538C>T)，SLC26A4(1174A>T、1226G>A、1229C>T、IVS7-2A>G、2168A>G)及线粒体MTRNR1(1494C>T、1555A>G)。同时，所有DNA样本均采用金标准Sanger测序法进行对比验证。本研究符合2013年修订的《世界医学协会赫尔辛基宣言》，并征得受试者知情同意。

①本研究来自250例受试者的250例DNA样本中，耳聋相关基因突变检出率为51.6%(129/250)，其中GJB2基因突变检出率为28.4%(71/250)，SLC26A4基因突变为19.2%(48/250)，线粒体MTRNR1基因突变为4.4%(11/250)，GJB3基因突变为2.0%(5/250)；129例检出的耳聋相关基因突变中，符合遗传病致病特征的耳聋相关基因突变的几率为86.8%(112/129)。②PCR-RDB法与Sanger测序法，对12个耳聋相关基因热点突变的检测结果均相同。③PCR-RDB法检测12个耳聋相关基因热点突变的灵敏度、特异度、总一致性、阳性预测值及阴性预测值均为100.0%。

虽然PCR-RDB法检测耳聋相关基因突变的结果与金标准Sanger测序法一致，但是本研究样本量尚小，因此是否可满足临床对遗传性耳聋基因检测的需求，则需多中心、大样本随机对照试验进一步研究、证实。

To verify the accuracy and effect of polymerase chain reaction-reverse dot blotting (PCR-RDB) technique in genetic deafness associated gene test.

A total of 250 patients with nonsyndromic hearing impairment(NSHI) who were diagnosed in Guangdong Women and Children Hospital from January 2014 to October 2016, were chosen as research objects. With PCR-RDB technique, we detected several genes associated with NSHI in genome DNA from peripheral blood cells of all the patients. To carry out PCR-RDB, the gene chip kit for hereditary hearing loss was used, which can detect 12 hot spots mutation of four genes in Chinese, including GJB2 (35del G, 176_191del 16, 235del C, 299_300del AT), GJB3 (538C>T), SLC26A4 (1174A>T, 1226G>A, 1229C>T, IVS7-2A>G, 2168A>G) and mitochondrion MTRNR1 (1494C>T, 1555A>G). Meanwhile, all the samples were also tested by Sanger sequencing, in order to compare and validate the results to that of PCR-RDB technique. This study was in line with the World Medical Association Declaration of Helsinki revised in 2013 and informed contents were obtained from all patients.

①There was 51.6%(129/250) genetic deafness associated gene mutations in DNA sample of 250 patients of NSHI, and some of which were 28.4%(71/250) from GJB2 gene, 19.2%(48/250) from SLC26A4 gene, 4.4%(11/250) from mitochondrion MTRNR1 gene and 2.0%(5/250) from GJB3 gene, respectively. Among 129 cases of deafness associated gene mutations, 86.8%(112/129) was met with pathogenicity characteristics of hereditary disease. ② All detection results of 12 hot spots mutations of deafness associated gene by PCR-RDB were consistent with those of by Sanger sequencing method. ③The sensitivity, specificity, total consistency, positive predictive value and negative predictive value of detection results of 12 hot spots mutations of deafness associated gene by PCR-RDB technique were all 100.0%.

Although detection results of gene hot spots mutation in genetic deafness associated gene by PCR-RDB technique were consistent with those of by Sanger sequencing method, whether it is worthy of clinical application, multicenter and large sample randomized controlled trial should be conducted as sample size of this study is still small.