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目的探讨在广泛开展的新生儿听力筛查中进行聋病基因同步筛查的可行性和实施方案与策略,以弥补新生儿听力筛查的不足和局限,并倡导在新生儿听力筛查中注入基因筛查的理念。方法2006年12月至2007年4月出生的460例新生儿作为研究对象,进行新生儿听力和基因的同步筛查。听力初步筛查采用筛查型耳声发射(otoacoustic emission,OAE),复筛采用筛查型 OAE结合自动判别听性脑干诱发电位(auto-auditory brainstem response,AABR)。在新生儿出生时或出生后3d 内采用自行设计的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脐带血或足跟血。遗传疾病筛查采样卡可以直接用于线粒体12SrRNA、GJB2基因、SLC26A4基因聚合酶链扩增反应(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。结果 460例新生儿听力初步筛查结果显示左耳未通过者9例,右耳未通过者3例,双耳均未通过者7例。经42d复筛后,初筛未通过19例中16例 OAE 和 AABR 均通过;1例右耳 OAE 未通过而 AABR 通过,左耳通过。2例患儿家长诉孩子听力正常而未来复筛。基因检测结果显示,460例新生儿中线粒体12SrRNAA1555G 酶切阳性者5例,经测序验证1例为12SrRNA A1555G 突变,3例为 C1556T 突变,1例测序结果显示正常序列;SLC26A4基因筛查 IVS7-2A>G 突变位点所在区域,发现5例 IVS7-2A>G 杂合携带者和1例 IVS7-18T>G 携带以及1例 IVS6-62_63insGT 携带者;GJB2基因筛查,发现8例235delC杂合携带者,4例 G109A 杂合携带者。共发现23例新生儿存在基因改变,其中1例为线粒体 A1555G致病突变,13例为致病基因突变的携带者,9例为基因多态改变。1例 A1555G 突变的新生儿及13例3个不同基因的携带者在新生儿听力初筛中均为通过。结论将听力筛查和基因筛查联合应用于早期发现处于语前听力损失或迟发型高危患儿或者是致聋基因的携带者,并结合定期的随诊及监测,是目前最为有力的筛查策略。倡导进行广泛的新生儿听力和基因的同步筛查工作。
OBJECTIVE: To investigate the feasibility and implementation of synchronous screening of deafness genes in the screening of neonatal hearing in a wide range of ways to make up for the shortcomings and limitations of neonatal hearing screening and to advocate the injection of neonatal hearing screening The concept of genetic screening. Methods A total of 460 newborns born from December 2006 to April 2007 were enrolled in the study. Initial hearing screening was performed with otoacoustic emission (OAE) screening. Screening OAE was used to screen for auto-auditory brainstem response (AABR). Neonatal umbilical cord blood or heel blood is collected at neonatal birth or within 3 days after birth by using a self-designed neonatal genetic screening screening card containing hearing screening information and blood sample information. Genetic disease screening sampling card can be directly used for mitochondrial 12SrRNA, GJB2 gene, SLC26A4 gene polymerase chain reaction (PCR). Alw26I restriction endonuclease screening of mitochondrial 12SrRNA A1555G point mutation, digested positive cases PCR product sequencing validation. The direct sequencing of PCR products was performed on the region of the GJB2 gene coding region and the region of the IVS7-2A> G mutation site of the SLC26A4 gene. DNAStar software for sequencing analysis of the results. Results 460 cases of neonatal hearing screening results showed that 9 cases did not pass the left ear, the right ear did not pass in 3 cases, none of the ears were in 7 cases. After screening for 42 days, 16 cases of OAE and AABR did not pass the primary screening in 19 cases, while one case of OAE failed in the right ear and AABR passed in the left ear. Two parents of children complained of hearing children normal and the future re-screening. Gene sequencing showed that mitochondrial 12SrRNAA1555G was positive in 5 out of 460 newborns. One case of 12SrRNA A1555G mutation was confirmed by sequencing and 3 cases of C1556T mutation. One case of sequencing showed normal sequence. SLC26A4 gene screening of IVS7-2A > G mutation sites found in 5 cases of IVS7-2A> G heterozygous carriers and 1 case of IVS7-18T> G carriers and 1 case of IVS6-62_63insGT carriers; GJB2 gene screening found 8 cases 235delC heterozygous carriers 4 cases of G109A heterozygous carriers. A total of 23 newborns were found to have gene mutations, of which 1 was mitochondrial A1555G pathogenic mutation, 13 were carriers of pathogenic mutations and 9 were polymorphic. One case of A1555G mutation in the newborn and 13 cases of three different gene carriers in neonatal hearing screening were passed. Conclusions Hearing screening and gene screening combined with early detection of carriers of prelingual or delayed-onset high-risk or deafness genes combined with regular follow-up and monitoring are the most powerful screening Strategy. Advocates for a wide range of neonatal hearing and genetic screening simultaneously.