基因芯片联合焦磷酸测序技术应用于新生儿遗传性聋基因突变位点筛查

Application of gene chip joint pyrosequencing technology in the newborn genetic deafness gene mutation screening

目的联合基因芯片和焦磷酸测序筛查新生儿遗传性聋基因突变,为早期诊断和预防遗传性聋提供理论依据。方法采集2000例新生儿抗凝脐带血,提取基因组DNA,采用微阵列芯片检测4个聋病基因共9个突变位点,对基因芯片检测的阳性结果进行焦磷酸测序验证。结果检出GJB2基因突变中有1例35delG突变类型,3例176 del16突变类型,57例235del C突变类型,9例299 del AT突变类型;6例GJB3基因538C>T突变类型;线粒体12S rRNA中有5例1555A>G突变类型,1例1494C>T突变类型;SLC26A4中有6例2168A>G突变类型,23例IVS7-2A>G突变类型。2000例新生儿中共103例携带突变基因,基因突变率5.15%。结论本地区非遗传性聋家族史新生儿中GJB2基因突变多见。新生儿中开展遗传性聋基因筛查对早期遗传性聋诊断和预防有非常重要的指导意义,尤其对线粒体12S rRNA基因突变诊断,能够预防用药控制聋病发生,保障该基因突变携带者健康具有十分重要意义。

OBJECTIVE To study the gene chipjoint pyrosequencing technology in the newborn genetic deafness gene mutation screening, and provide a theoretical basis for the early diagnosis and prevention of genetic deafness. METHODS 2000 Neonatal EDTA umbilical cord blood was collected and genomic DNA （gDNA） was extracted. Microarray chip was used to detect four deafness gene at 9 mutation sites. And the positive result of gene chip detection was verified by pyrosequeneing. RESULTS Among the GJB2 mutations, there were 1 case of 35delG mutation type, 3 cases of 176 dell6 mutation type, 57 eases of 235del C mutation type, 9 cases of 299 del AT mutation type, 6 cases of GJB3 gene 538C〉T mutation type. There were 5 cases of 1555A〉G mutations and 1 case of 1494C〉T mutations in mitochondrial 12S rRNA. There were 6 eases of 2168A〉G mutation type and 23 cases of IVST-2A〉G mutations in SLC26A4. 103 cases of newborns carry the mutated gene in 2,000, the gene mutation rate is 5.15%. CONCLUSION All the four genes mutation at nine mutation sites are found in newborns with family history of non-hereditary deafness, and GJB2 gene mutation is common. The screening of genetic deafness in newborns is very important for early diagnosis and prevention of hereditary hearing loss. In particular, the diagnosis of mitochondrial 12S rRNA gene mutation can prevent the occurrence of deafness caused by drug use, for the genetic mutation of these carriers＇ health is of great significance.