基因芯片联合二代测序技术在产前耳聋基因筛查中的应用价值
|
摘要
目的探究基因芯片(GC)联合二代测序技术(NGS)检测在产前耳聋基因筛查中的应用价值。方法选取2016年1月到2018年1月在我院产前筛查中心行孕妇外周血耳聋基因芯片及二代测序技术检测的孕妇及其新生儿资料。使用自身对照法,回顾性分析GC、NGS筛查和两者联合使用三种方法之间的准确率、灵敏度以及特异度等指标。结果全部病例420例,有效随访者360例,最终确诊为携带耳聋突变基因孕妇为18例,发病率为5.0%。其中GJB2基因突变11例(包括6例235delC和5例299-300delAT位点突变),携带率为3.1%(11/360);GJB3基因突变3例(包括2例538C>T和1例1555A>G位点突变),携带率为0.8%(3/360);SLC26A4(PDS基因突变4例(均为IVS7-2A>G位点突变),携带率为1.1%(4/360)。结论 GC联合NGS法应用于产前耳聋基因的筛查,可提高其诊断准确率和特异度,具有较高的临床应用价值,值得进一步推广使用。
Objective:Explore the value of gene chip(GC)combined with second-generation sequencing technology(NGS)in the screening of prenatal deafness genes. Methods:The data of pregnant women and their newborns were collected from the pregnant women′s peripheral blood lobe gene chip and the second generation sequencing technology from January 2016 to January 2018 in our prenatal screening center. Using self-control methods,retrospective analysis of GC,NGS screening,and the combination of the three methods used in the three methods of accuracy,sensitivity and specificity. Results:There were 420 cases in all cases and 360 cases were effectively followed up. The final diagnosis was 18 cases of pregnant women with deafness mutation gene,the incidence rate was 5.0%. Among them,11 cases of GJB2 gene mutation(including 6 cases of 235 delC and 5 cases of 299-300 delAT site mutation),the carrying rate was 3.1%(11/360);3 cases of GJB3 gene mutation(including 2 cases of 538 C>T and 1 case of 1555 A>G site mutation),carrying rate of 0.8%(3/360);SLC26 A4(PDS gene mutation in 4 cases(all IVS7-2 A>G site mutation),the carrying rate was 1.1%(4/360). Conclusion:The GC combined with NGS method is applied to the screening of prenatal deafness genes,which can improve the diagnostic accuracy and specificity. It has high clinical application value and is worthy of further promotion.
Objective:Explore the value of gene chip(GC)combined with second-generation sequencing technology(NGS)in the screening of prenatal deafness genes. Methods:The data of pregnant women and their newborns were collected from the pregnant women′s peripheral blood lobe gene chip and the second generation sequencing technology from January 2016 to January 2018 in our prenatal screening center. Using self-control methods,retrospective analysis of GC,NGS screening,and the combination of the three methods used in the three methods of accuracy,sensitivity and specificity. Results:There were 420 cases in all cases and 360 cases were effectively followed up. The final diagnosis was 18 cases of pregnant women with deafness mutation gene,the incidence rate was 5.0%. Among them,11 cases of GJB2 gene mutation(including 6 cases of 235 delC and 5 cases of 299-300 delAT site mutation),the carrying rate was 3.1%(11/360);3 cases of GJB3 gene mutation(including 2 cases of 538 C>T and 1 case of 1555 A>G site mutation),carrying rate of 0.8%(3/360);SLC26 A4(PDS gene mutation in 4 cases(all IVS7-2 A>G site mutation),the carrying rate was 1.1%(4/360). Conclusion:The GC combined with NGS method is applied to the screening of prenatal deafness genes,which can improve the diagnostic accuracy and specificity. It has high clinical application value and is worthy of further promotion.
引文
[1]新生儿聋病易感基因筛查的研究进展[J].听力学及言语疾病杂志,2015(1):91-96.
[2]凌寒.卫生部发布《中国出生缺陷防治报告(2012)》[J].中国药房,2012(39):1-1.
[3]Paz-Y-Mi?o C,Beaty D,López-Cortés A,et al.Frequency of GJB2,and del(GJB6-D13S1830)mutations among an Ecuadorian mestizo population[J].International Journal of Pediatric Otorhinolaryngology,2014,78(10):1648-1654.
[4]Pan J,Xu P,Tang W,et al.Mutation analysis of common GJB2,SCL26A4 and 12S rRNA genes among 380 deafness patients in northern China.[J].International Journal of Pediatric Otorhinolaryngology,2017,98:39.
[5]Estivill X,Fortina P,Surrey S,et al.Connexin-26 mutations in sporadic and inherited sensorineural deafness.[J].Lancet,2016,351(9109):394-8.
[6]张昊昱,张宁,张华,等.耳聋基因检测在遗传性耳聋诊断及遗传咨询中的应用[J].中华耳科学杂志,2016,14(5):639-643.
[7]Boland P M,Ruth K,Matro J M,et al.Genetic counselors′(GC)knowledge,awareness,and understanding of clinical next-generation sequencing(NGS)genomic testing[J].Clinical Genetics,2015,88(6):565-572.
[8]Ma Y,Xiao Y,Bai X,et al.GJB2,SLC26A4,and mitochondrial DNA12S rRNA hot-spots in 156 subjects with non-syndromic hearing loss in Tengzhou,China[J].Acta oto-laryngologica,2016,136(8):1.
[9]柴福,赵海亮,邱书奇.新生儿GJB2、GJB3、SLC26A4和线粒体DNA12SrRNA基因突变筛查结果分析[J].临床耳鼻咽喉头颈外科杂志,2017(9):664-666.
[10]Dingyuan M A,Zhang J,Luo C,et al.Genetic counseling for patients with nonsyndromic hearing impairment directed by gene analysis:[J].Molecular Medicine Reports,2016,13(3):1967-1974.
[11]Young K S,Reum K A,Kim N K D,et al.Unraveling of Enigmatic Hearing-ImpairedGJB2Single Heterozygotes by Massive Parallel Sequencing:DFNB1 or Not?:[J].Medicine,2016,95(14):e3029.
[12]李庆忠,王秋菊,赵立东,等.国人非综合征型遗传性聋患者GJB3基因突变分析[J].听力学及言语疾病杂志,2005,13(3):145-148.
[13]Koch Y,Van B F,Kaiser S,et al.Connexin 31(GJB3)deficiency in mouse trophoblast stem cells alters giant cell differentiation and leads to loss of oxygen sensing.[J].Biology of Reproduction,2012,87(2):37.
[14]牛志杰,冯永,梅凌云,等.非综合征型X连锁隐性遗传耳聋家系临床表型及遗传学特征分析[J].中华耳科学杂志,2017,15(2):195-200.
[15]Tong G X,Chang Q,Hamele-Bena D,et al.Targeted NextGeneration Sequencing Analysis of a Pendred SyndromeAssociated Thyroid Carcinoma[J].Endocrine Pathology,2016,27(1):70-75.
[16]Pandey N,Rashid T,Jalvi R,et al.Mutations in OTOF,CLDN14&SLC26A4 genes as major causes of hearing impairment in Dhadkai village,Jammu&Kashmir,India.[J].Indian Journal of Medical Research,2017,146(4):489-497.
[17]Li X,Sanneman J D,Harbidge D G,et al.SLC26A4 Targeted to the Endolymphatic Sac Rescues Hearing and Balance in Slc26a4Mutant Mice[J].Plos Genetics,2016,9(7):e1003641.
[2]凌寒.卫生部发布《中国出生缺陷防治报告(2012)》[J].中国药房,2012(39):1-1.
[3]Paz-Y-Mi?o C,Beaty D,López-Cortés A,et al.Frequency of GJB2,and del(GJB6-D13S1830)mutations among an Ecuadorian mestizo population[J].International Journal of Pediatric Otorhinolaryngology,2014,78(10):1648-1654.
[4]Pan J,Xu P,Tang W,et al.Mutation analysis of common GJB2,SCL26A4 and 12S rRNA genes among 380 deafness patients in northern China.[J].International Journal of Pediatric Otorhinolaryngology,2017,98:39.
[5]Estivill X,Fortina P,Surrey S,et al.Connexin-26 mutations in sporadic and inherited sensorineural deafness.[J].Lancet,2016,351(9109):394-8.
[6]张昊昱,张宁,张华,等.耳聋基因检测在遗传性耳聋诊断及遗传咨询中的应用[J].中华耳科学杂志,2016,14(5):639-643.
[7]Boland P M,Ruth K,Matro J M,et al.Genetic counselors′(GC)knowledge,awareness,and understanding of clinical next-generation sequencing(NGS)genomic testing[J].Clinical Genetics,2015,88(6):565-572.
[8]Ma Y,Xiao Y,Bai X,et al.GJB2,SLC26A4,and mitochondrial DNA12S rRNA hot-spots in 156 subjects with non-syndromic hearing loss in Tengzhou,China[J].Acta oto-laryngologica,2016,136(8):1.
[9]柴福,赵海亮,邱书奇.新生儿GJB2、GJB3、SLC26A4和线粒体DNA12SrRNA基因突变筛查结果分析[J].临床耳鼻咽喉头颈外科杂志,2017(9):664-666.
[10]Dingyuan M A,Zhang J,Luo C,et al.Genetic counseling for patients with nonsyndromic hearing impairment directed by gene analysis:[J].Molecular Medicine Reports,2016,13(3):1967-1974.
[11]Young K S,Reum K A,Kim N K D,et al.Unraveling of Enigmatic Hearing-ImpairedGJB2Single Heterozygotes by Massive Parallel Sequencing:DFNB1 or Not?:[J].Medicine,2016,95(14):e3029.
[12]李庆忠,王秋菊,赵立东,等.国人非综合征型遗传性聋患者GJB3基因突变分析[J].听力学及言语疾病杂志,2005,13(3):145-148.
[13]Koch Y,Van B F,Kaiser S,et al.Connexin 31(GJB3)deficiency in mouse trophoblast stem cells alters giant cell differentiation and leads to loss of oxygen sensing.[J].Biology of Reproduction,2012,87(2):37.
[14]牛志杰,冯永,梅凌云,等.非综合征型X连锁隐性遗传耳聋家系临床表型及遗传学特征分析[J].中华耳科学杂志,2017,15(2):195-200.
[15]Tong G X,Chang Q,Hamele-Bena D,et al.Targeted NextGeneration Sequencing Analysis of a Pendred SyndromeAssociated Thyroid Carcinoma[J].Endocrine Pathology,2016,27(1):70-75.
[16]Pandey N,Rashid T,Jalvi R,et al.Mutations in OTOF,CLDN14&SLC26A4 genes as major causes of hearing impairment in Dhadkai village,Jammu&Kashmir,India.[J].Indian Journal of Medical Research,2017,146(4):489-497.
[17]Li X,Sanneman J D,Harbidge D G,et al.SLC26A4 Targeted to the Endolymphatic Sac Rescues Hearing and Balance in Slc26a4Mutant Mice[J].Plos Genetics,2016,9(7):e1003641.