新生儿聋病基因（GJB2、SLC26A4、线粒体12SrRNA）的分子流行病学研究
|
摘要
目的
通过对新生儿开展聋病易感基因筛查,即对新生儿(GJB2、SLC26A4、线粒体12SrRNA)基因突变情况进行检测,探讨新生儿中聋病易感基因的分子流行病学特点,为制定防聋治聋策略提供依据。
方法
以2008年1月至2009年6月出生于中国医科大学附属盛京医院产科病房的659例新生儿作为研究对象。在新生儿生后3天,采用含有听力筛查信息和血样信息的新生儿遗传疾病筛查采样卡,采集新生儿点滴微量足跟血(不超过125ul)。采样卡可以直接用于线粒体12SrRNA、GJB2基因、SLC26A4基因的聚合酶链扩增反应(polymerase chain reaction,PCR)。Alw26I限制性内切酶筛查线粒体12SrRNA A1555G点突变,对酶切阳性病例进行PCR产物测序验证。对GJB2基因编码区和SLC26A4基因IVS7-2A>G突变位点所在区域进行PCR产物的直接测序。DNAStar软件对测序结果进行比对分析。
对基因突变的新生儿进行流行病学特点分析。
结果
659例新生儿中GJB2基因235delC杂合突变7例,致病突变的携带率为1.06%; SLC26A4基因IVS7-2A>G杂合突变3例,致病突变的携带率为0.46%,三项基因总突变率1.52%；携带突变基因的新生儿男性占2.01%,女性占0.96%；满族新生儿致病突变的携带率最高,为1.82%,汉族为1.57%。
结论
结论659例新生儿中耳聋易感基因突变率为1.52%,有男性多于女性、满族多于汉族的趋势。在新生儿中开展聋病易感基因筛查,早期发现遗传性耳聋高危人群,是降低耳聋发病率、提高人口素质的重要措施。
Objective
To discuss the report of newborn gene screening in order to find the feature of the epidemiologic of the pathogenic in Liaoning province.discuss the relationship of the genes mutations and facters. providing a basis to draft pre-deaf and treat-deaf strategy.
Method
six hundred and fiftynine newborn babies from Feb.2008 to Jun 2009 accepted the gene screening. Newborn genetic disease screening cards (the utility Model Patent-200720103139.4) were used for collecting the blood spot from the calcar pedis within the moment of newborn. The cards can be directly performed the polymerase chain reaction (PCR) for screening the mitochondrial 12SrRNA 1555G and GJB2 as well as SLC26A4 genes mutations. The restriction enzyme Alw26I was used to recognize the point mutation of 12SrRNA A1555G. The samples with the possible 12SrRNA A1555G mutation were then sequenced to verify. The PCR products from the GJB2 coding region and SLC26A4 IVS7-2A>G hot spot region were sequenced directly. The software of DNAStar was used to analysis the sequence,
molecular epidemiology discussion of the newborn with genes mutations.
Result
For the GJB2 gene screening, seven were 235delC heterozygote,1.06%; For the SLC26A4 gene screening, three with the heterozygote of IVS7-2A>G mutation was found,0.46%; Totally 1.52%of newborn carried the genes mutaion.2.01% male carried the genes mutation,female 0.96%.1.82% of the Manchu carried the genes mutation,Han 1.57%.
Conclusion
It may be all the gene screening found 10 newborn babies of the 659 harbored the changes in the three genes. Totally 1.52%of newborn carried the genes mutaion.We found the tendency is male has more carried mutation than female,Manchu has more carried mutation than Han.Newborn gene screening for the purpose of early diagnosis and discovery the prelingual or late-onset or the high risk as well as the pathogenic carriers.This providing a basis to draft pre-deaf and treat-deaf strategy.
通过对新生儿开展聋病易感基因筛查,即对新生儿(GJB2、SLC26A4、线粒体12SrRNA)基因突变情况进行检测,探讨新生儿中聋病易感基因的分子流行病学特点,为制定防聋治聋策略提供依据。
方法
以2008年1月至2009年6月出生于中国医科大学附属盛京医院产科病房的659例新生儿作为研究对象。在新生儿生后3天,采用含有听力筛查信息和血样信息的新生儿遗传疾病筛查采样卡,采集新生儿点滴微量足跟血(不超过125ul)。采样卡可以直接用于线粒体12SrRNA、GJB2基因、SLC26A4基因的聚合酶链扩增反应(polymerase chain reaction,PCR)。Alw26I限制性内切酶筛查线粒体12SrRNA A1555G点突变,对酶切阳性病例进行PCR产物测序验证。对GJB2基因编码区和SLC26A4基因IVS7-2A>G突变位点所在区域进行PCR产物的直接测序。DNAStar软件对测序结果进行比对分析。
对基因突变的新生儿进行流行病学特点分析。
结果
659例新生儿中GJB2基因235delC杂合突变7例,致病突变的携带率为1.06%; SLC26A4基因IVS7-2A>G杂合突变3例,致病突变的携带率为0.46%,三项基因总突变率1.52%；携带突变基因的新生儿男性占2.01%,女性占0.96%；满族新生儿致病突变的携带率最高,为1.82%,汉族为1.57%。
结论
结论659例新生儿中耳聋易感基因突变率为1.52%,有男性多于女性、满族多于汉族的趋势。在新生儿中开展聋病易感基因筛查,早期发现遗传性耳聋高危人群,是降低耳聋发病率、提高人口素质的重要措施。
Objective
To discuss the report of newborn gene screening in order to find the feature of the epidemiologic of the pathogenic in Liaoning province.discuss the relationship of the genes mutations and facters. providing a basis to draft pre-deaf and treat-deaf strategy.
Method
six hundred and fiftynine newborn babies from Feb.2008 to Jun 2009 accepted the gene screening. Newborn genetic disease screening cards (the utility Model Patent-200720103139.4) were used for collecting the blood spot from the calcar pedis within the moment of newborn. The cards can be directly performed the polymerase chain reaction (PCR) for screening the mitochondrial 12SrRNA 1555G and GJB2 as well as SLC26A4 genes mutations. The restriction enzyme Alw26I was used to recognize the point mutation of 12SrRNA A1555G. The samples with the possible 12SrRNA A1555G mutation were then sequenced to verify. The PCR products from the GJB2 coding region and SLC26A4 IVS7-2A>G hot spot region were sequenced directly. The software of DNAStar was used to analysis the sequence,
molecular epidemiology discussion of the newborn with genes mutations.
Result
For the GJB2 gene screening, seven were 235delC heterozygote,1.06%; For the SLC26A4 gene screening, three with the heterozygote of IVS7-2A>G mutation was found,0.46%; Totally 1.52%of newborn carried the genes mutaion.2.01% male carried the genes mutation,female 0.96%.1.82% of the Manchu carried the genes mutation,Han 1.57%.
Conclusion
It may be all the gene screening found 10 newborn babies of the 659 harbored the changes in the three genes. Totally 1.52%of newborn carried the genes mutaion.We found the tendency is male has more carried mutation than female,Manchu has more carried mutation than Han.Newborn gene screening for the purpose of early diagnosis and discovery the prelingual or late-onset or the high risk as well as the pathogenic carriers.This providing a basis to draft pre-deaf and treat-deaf strategy.
引文
1 刘学忠,欧阳小梅,Van D,et al中国人群遗传性耳聋研究进展.中华耳科学杂志,2006,4(2)：81-89.
2 Morton NE,Genetic epidemiology of hearing impairment.Ann N Y Acad Sci,1991,630:16-31.
3 Tay JP,Metcalfe RA,Watson PF,et al.Mutations of the PDS gene,encoding pendrin, are associated with protein mislocalization and loss of iodide efflux:implications for thyroid dysfunction in Pendred syndrome[J]. J Clin Endocrinol Metab,2002,87:1778.
4 Wang QJ, Zhao YL,Lan L,et al. A distinct spectrum of SLC26A4 mutations in patients with enlarged vestibular aqueduct in China[J].Clinical Genetics,2007,72:245.
5 王秋菊,新生儿聋病基因筛查—悄然的革命.听力学及言语疾病杂志,2008,16(2)83-88.
6 SmithRJH, RobinNH。Genetic testing for deafness-GJB2 and SLC26A4 as causes of deafness.J Communication Disorders,2002,35:367-377.
7 Yuan HJ, Qian YP, Xu YJ, Cao J, Bai L, Shen W, Ji F, Zhang X,Kang D, Mo JQ, Grienwald JH, Han D, Zhai S, Young WY, GuanMX. Cosegregation of the G7444A mutation in the mitochondrialCOI/tRNA Ser(UCN) gene with the 12S rRNA A1555G mutation in aChinese family with aminoglycoside-induced and nonsyndromichearing loss. Am J Med Genet A, 2005,138(2):133-140.
8 赵亚丽,王秋菊,翟所强.前庭水管扩大与SLC26A4基因分子流行病学研究.中国人民解放军总医院硕士学位毕业论文,2006.
9 李琦,戴朴,黄德亮,等.SLC26A4基因IVS7-2A>G突变在中国不同地区和民族重度感音性聋患者中分布频率的观察.中华耳鼻咽喉头颈外科杂志.2007.12：893-897.
10 王秋菊.新生儿聋病基因筛查实施方案与策略研究.中华耳鼻咽喉头颈外科杂志.2007.11：809-813.
11 王秋菊.新生儿聋病易感基因筛的意义与策略.中国医学文摘耳鼻咽喉科学.2007.22：21-22.
1 刘学忠,欧阳小梅,Yan D,et al.中国人群遗传性耳聋研究进展.中华耳科学杂志,2006,4(2)：81-89.
2 Morton NE,Genetic epidemiology of hearing impairment,Ann N Y Acad Sci,1991,630:16-31.
3 王国建,戴扑,韩东一,基因芯片技术在非综合征性耳聋快速基因诊断中的应用研究.中华耳科学杂志,2008(6)1：61-66.
4 徐志勇,王沙燕,GJB2基因突变与临床表型关系的研究进展.中国优生与遗传杂志,2008,16(8)3-4.
5 Forge A,Becker D,Casalotti S,et al.Gap junctions in inner ear:comparison of distribution patterns in different vertebrates and assessement of connexin composition in mammals[J].J Comp Neurol,2003,467:207.
6 Kudo T,ikeda K,KureS,et al Novelmutations in connexin 26gene (GJB2)responsible for childhood deafness in the Japanese population [J].Am J Med Genet,2000,90:141-145
7 李建瑞,陈瑛,郭皖北,等.中国人GJB2耳聋基因突变分析[J].中华医学遗传学杂志,2003,20(5)：441-443
8 郭玉芬,刘晓雯,关静,等.西北地区非综合征型耳聋患者GJB2、SLC26A4基因突变的分子流行病学研究.听力学及言语疾病杂志,2008,16(4)363-266
9 Gualandi F, Ravani A,Berto A,et al Exploring the clinical and epidemiological complexity of GJB2 linked deafness [J]. Am J Med Genet,2002,112:38-45.
10 Loffler J, Nekahm D, Hirst-stalmann A,et al Sensorineural hearing loss and incidence of Cx26 mutations in Austria[J]. Eur J Hum Genet,2001,9:226-230
11 Green GE, Scott DA,McDonald JM,et al Carrier rates in the midwestem United States for GJB2 mutations causing inherited deafness[J].JAMA,1999,281:2211-2216
12 Tay JP,Metcalfe RA,Watson PF,et al Mutations of the PDS gene,encoding pendrin, are associated with protein mislocalization and loss of iodide efflux:implications for thyroid dysfunction in Pendred syndrome[J]. J Clin Endocrinol Metab,2002,87:1778.
13 Wang QJ, Zhao YL,Lan L,et al. A distinct spectrum of SLC26A4 mutations in patients with enlarged vestibular aqueduct in China[J].Clinical Genetics,2007,72:245
14 王秋菊.新生儿聋病基因筛查——悄然的革命.听力学及言语疾病杂志,2008,16(2)83-88
15 Yuan HJ, Qian YP, Xu YJ, Cao J, Bai L, Shen W, Ji F, Zhang X,Kang D, Mo JQ, Grienwald JH, Han D, Zhai S, Young WY, GuanMX. Cosegregation of the G7444A mutation in the mitochondrialCOI/tRNA Ser(UCN) gene with the 12S rRNA A1555G mutation in aChinese family with aminoglycoside-induced and nonsyndromichearing loss. Am J Med Genet A, 2005,138(2):133-140.
16 Masmoudi S. Elgaied-Boulila A, Kassab I, et al. Determination of the frequency of connexin26 mutations in inherited sensorineural deafness andcarrier rates in the Tunisian population using DGGE.J Med Genet,2000,37:E39.
17 Liu XZ.Xia XJ, Ke XM,et al. The prevalence of connexin 26 (GJB2) mutations in the Chinese population. Hum Genet,2002,111:394-397.
18戴朴,杨伟炎,韩东一,等,Prev-DAF试剂盒分析线粒体基因1555A-G突变中华耳科学杂志，2004，2：37-411
19戴朴,韩东一,杨伟炎,标准聋病分子诊断实验室的结构和功能。中华耳科学杂志。2004,2：60-63。
20 SmithRJH, RobinNH。Genetic testing for deafness-GJB2 and SLC26A4 as causes of deafness.J Communication Disorders,2002,35:367-377.
2 Morton NE,Genetic epidemiology of hearing impairment.Ann N Y Acad Sci,1991,630:16-31.
3 Tay JP,Metcalfe RA,Watson PF,et al.Mutations of the PDS gene,encoding pendrin, are associated with protein mislocalization and loss of iodide efflux:implications for thyroid dysfunction in Pendred syndrome[J]. J Clin Endocrinol Metab,2002,87:1778.
4 Wang QJ, Zhao YL,Lan L,et al. A distinct spectrum of SLC26A4 mutations in patients with enlarged vestibular aqueduct in China[J].Clinical Genetics,2007,72:245.
5 王秋菊,新生儿聋病基因筛查—悄然的革命.听力学及言语疾病杂志,2008,16(2)83-88.
6 SmithRJH, RobinNH。Genetic testing for deafness-GJB2 and SLC26A4 as causes of deafness.J Communication Disorders,2002,35:367-377.
7 Yuan HJ, Qian YP, Xu YJ, Cao J, Bai L, Shen W, Ji F, Zhang X,Kang D, Mo JQ, Grienwald JH, Han D, Zhai S, Young WY, GuanMX. Cosegregation of the G7444A mutation in the mitochondrialCOI/tRNA Ser(UCN) gene with the 12S rRNA A1555G mutation in aChinese family with aminoglycoside-induced and nonsyndromichearing loss. Am J Med Genet A, 2005,138(2):133-140.
8 赵亚丽,王秋菊,翟所强.前庭水管扩大与SLC26A4基因分子流行病学研究.中国人民解放军总医院硕士学位毕业论文,2006.
9 李琦,戴朴,黄德亮,等.SLC26A4基因IVS7-2A>G突变在中国不同地区和民族重度感音性聋患者中分布频率的观察.中华耳鼻咽喉头颈外科杂志.2007.12：893-897.
10 王秋菊.新生儿聋病基因筛查实施方案与策略研究.中华耳鼻咽喉头颈外科杂志.2007.11：809-813.
11 王秋菊.新生儿聋病易感基因筛的意义与策略.中国医学文摘耳鼻咽喉科学.2007.22：21-22.
1 刘学忠,欧阳小梅,Yan D,et al.中国人群遗传性耳聋研究进展.中华耳科学杂志,2006,4(2)：81-89.
2 Morton NE,Genetic epidemiology of hearing impairment,Ann N Y Acad Sci,1991,630:16-31.
3 王国建,戴扑,韩东一,基因芯片技术在非综合征性耳聋快速基因诊断中的应用研究.中华耳科学杂志,2008(6)1：61-66.
4 徐志勇,王沙燕,GJB2基因突变与临床表型关系的研究进展.中国优生与遗传杂志,2008,16(8)3-4.
5 Forge A,Becker D,Casalotti S,et al.Gap junctions in inner ear:comparison of distribution patterns in different vertebrates and assessement of connexin composition in mammals[J].J Comp Neurol,2003,467:207.
6 Kudo T,ikeda K,KureS,et al Novelmutations in connexin 26gene (GJB2)responsible for childhood deafness in the Japanese population [J].Am J Med Genet,2000,90:141-145
7 李建瑞,陈瑛,郭皖北,等.中国人GJB2耳聋基因突变分析[J].中华医学遗传学杂志,2003,20(5)：441-443
8 郭玉芬,刘晓雯,关静,等.西北地区非综合征型耳聋患者GJB2、SLC26A4基因突变的分子流行病学研究.听力学及言语疾病杂志,2008,16(4)363-266
9 Gualandi F, Ravani A,Berto A,et al Exploring the clinical and epidemiological complexity of GJB2 linked deafness [J]. Am J Med Genet,2002,112:38-45.
10 Loffler J, Nekahm D, Hirst-stalmann A,et al Sensorineural hearing loss and incidence of Cx26 mutations in Austria[J]. Eur J Hum Genet,2001,9:226-230
11 Green GE, Scott DA,McDonald JM,et al Carrier rates in the midwestem United States for GJB2 mutations causing inherited deafness[J].JAMA,1999,281:2211-2216
12 Tay JP,Metcalfe RA,Watson PF,et al Mutations of the PDS gene,encoding pendrin, are associated with protein mislocalization and loss of iodide efflux:implications for thyroid dysfunction in Pendred syndrome[J]. J Clin Endocrinol Metab,2002,87:1778.
13 Wang QJ, Zhao YL,Lan L,et al. A distinct spectrum of SLC26A4 mutations in patients with enlarged vestibular aqueduct in China[J].Clinical Genetics,2007,72:245
14 王秋菊.新生儿聋病基因筛查——悄然的革命.听力学及言语疾病杂志,2008,16(2)83-88
15 Yuan HJ, Qian YP, Xu YJ, Cao J, Bai L, Shen W, Ji F, Zhang X,Kang D, Mo JQ, Grienwald JH, Han D, Zhai S, Young WY, GuanMX. Cosegregation of the G7444A mutation in the mitochondrialCOI/tRNA Ser(UCN) gene with the 12S rRNA A1555G mutation in aChinese family with aminoglycoside-induced and nonsyndromichearing loss. Am J Med Genet A, 2005,138(2):133-140.
16 Masmoudi S. Elgaied-Boulila A, Kassab I, et al. Determination of the frequency of connexin26 mutations in inherited sensorineural deafness andcarrier rates in the Tunisian population using DGGE.J Med Genet,2000,37:E39.
17 Liu XZ.Xia XJ, Ke XM,et al. The prevalence of connexin 26 (GJB2) mutations in the Chinese population. Hum Genet,2002,111:394-397.
18戴朴,杨伟炎,韩东一,等,Prev-DAF试剂盒分析线粒体基因1555A-G突变中华耳科学杂志，2004，2：37-411
19戴朴,韩东一,杨伟炎,标准聋病分子诊断实验室的结构和功能。中华耳科学杂志。2004,2：60-63。
20 SmithRJH, RobinNH。Genetic testing for deafness-GJB2 and SLC26A4 as causes of deafness.J Communication Disorders,2002,35:367-377.