儿童氨基糖苷类药物性耳聋基因快速无创检测法的应用

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摘要目的：探讨氨基糖苷类药物性耳聋相关基因位点A1555G和C1494T的快速无创检测法在儿童耳聋诊断中的应用。方法：选取2015年5月至2017年5月温州医科大学附属第一医院诊断为非综合征型感音神经性耳聋患儿126例，收集其口腔黏膜拭子及静脉血。采用多重等位基因特异性PCR（MAS-PCR）检测口腔拭子来源DNA中A1555G和C1494T突变情况。同时用Sanger测序法检测静脉血来源DNA中A1555G和C1494T突变情况，并通过Kappa一致性检验分析2种方法检出率的一致性来验证前者的可靠性。结果：采用MAS-PCR检测口腔黏膜拭子来源DNA中A1555G和C1494T突变的检出率为6.35%（8/126），其中A1555G突变7例、C1494T突变1例。静脉血Sanger测序法的检出率为6.35%（8/126），Kappa一致性检验结果显示2种方法检出率一致性较好（kappa=1，P＜0.01）。结论：采用MAS-PCR检测口腔黏膜拭子来源DNA中A1555G和C1494T突变是一种简便、快速、无创的检测手段，为婴幼儿，乃至新生儿药物性耳聋检测的开展提供一种可行的方法。

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	张琼敏
	李斯斯
	陈君
	朱翌

关键词 ：口腔黏膜拭子, 氨基糖苷类药物性耳聋, 多重等位基因特异性PCR, 无创

Abstract：Objective: To probe the application of a fast and noninvasive method for testing the mutations A1555G and C1494T associated with aminoglycoside-induced hearing loss in children. Methods: A total of 126 non-syndromic sensorineural deafness patients aged 2 to 15 were selected for collecting oral swabs and venous blood from the First Affiliated Hospital of Wenzhou Medical University during May 2015 to May 2017. Multiplex allele-specific PCR was used to test the mutations of A1555G and C1494T in DNA from oral swabs. Simultaneously, Sanger sequencing was used to detect the mutations of A1555G and C1494T in the DNA of vein blood to verify the reliability of the former by Kappa testing. Results: The detection rate of A1555G and C1494T mutations in DNA from oral swabs detected by multiplex allele-specific PCR was 6.35% (8/126), including 7 cases of A1555G mutations and 1 cases of C1494T mutation. The testing rate of Sanger sequencing was also 6.35% (8/126), and the two methods were in a good consistency checking (kappa=1, P<0.01). Conclusion: The testing of A1555G and C1494T mutations with multiplex allele-specific PCR in DNA from oral buccal swabs is a simple, rapid and non-invasive method to provide another option for the testing of aminoglycoside-induced deafness in infants and newborns.

Key words： oral buccal swab aminoglycoside-induced hearing loss multiplex allele-specific PCR non-invasive

收稿日期: 2018-05-26

基金资助:国家自然科学基金资助项目（81070794）；温州市公益性科技计划项目（Y20160416）。

通讯作者: 朱翌，主任医师，Email：269939306@qq.com。

作者简介: 张琼敏（1984-），女，浙江温州人，住院医师，硕士。

链接本文:

https://xb.wmu.edu.cn/CN/ 或 https://xb.wmu.edu.cn/CN/Y2018/V48/I11/791

[1] YOSHIKAWA S, KAWANO A, HAYASHI C, et al. The clinical features of patients with the homozygous 235delC and the compound-heterozygous Y136X/G45E of the GJB2 mutations (Connexin 26) in cochlear implant recipients[J]. Auris Nasus Larynx, 2011, 38(4): 444-449.
[2] 孙喜斌, 于丽玫, 曲成毅, 等. 中国听力残疾构成特点及康复对策[J]. 中国听力语言康复科学杂志, 2008, 27(2): 21-24.
[3] STEVENS G, FLAXMAN S, BRUNSKILL E, et al. Global and regional hearing impairment prevalence: an analysis of 42 studies in 29 countries[J]. Eur J Public Health, 2013, 23 (1): 146-152.
[4] 彭光华, 王辉, 姚娟, 等. 浙江省余姚地区22个非综合征型耳聋家系遗传分析[J]. 温州医科大学学报, 2016, 46(10): 709-715.
[5] MUTAI H, WATABE T, KOSAKI K, et al. Mitochondrial mutations in maternally inherited hearing loss[J]. BMC Med Genet, 2017, 18(1): 32.
[6] LU J, LI Z, ZHU Y, et al. Mitochondrial 12S rRNA variants in 1642 Han Chinese pediatric subjects with aminoglycoside-induced and nonsyndromic hearing loss[J]. Mitochondrion, 2010, 10(4): 380-390.
[7] GAO Z, CHEN Y, GUAN M X. Mitochondrial DNA mutations associated with aminoglycoside induced ototoxicity[J]. J Otol, 2017, 12(1): 1-8.
[8] 郑静, 杨爱芬, 张婷, 等. 多重等位基因PCR检测线粒体12S rRNA基因突变[J]. 中华检验医学杂志, 2011, 34(7): 628-632.
[9] 巫静帆, 李小霞, 谭淑娟, 等. 东莞户籍33 810例新生儿听力筛查联合耳聋基因检测与分析[J]. 中华耳科学杂志, 2018, 16(2): 176-180.
[10] FISCHEL-GHODSIAN N. Genetic factors in aminoglycoside toxicity[J]. Pharmacogenomics, 2005, 6(1): 27-36.
[11] 戴朴, 刘新, 于飞, 等. 18个省市聋校学生非综合征性聋病分子流行病学研究(I)——GJB2 235delC和线粒体DNA 12SrRNA A1555G突变筛查报告[J]. 中华耳科学杂志, 2006, 4(1): 1-5.
[12] QI L, PING L, XUE-JUN D, et al. A novel method for detection the deafness-associated mitochondrial A1555G and C1494T mutations[J]. Clin Lab, 2016, 62(3): 477-481.
[13] WANG L, WANG X, CAI X, et al. Study of mitochondrial DNA A1555G and C1494T mutations in a large cohort of women individuals[J]. Mitochondrial DNA A DNA Mapp Seq Anal, 2018: 1-4.
[14] MENG F, HE Z, TANG X, et al. Contribution of the tRNAIle 4317A→G mutation to the phenotypic manifestation of the deafness-associated mitochondrial 12S rRNA 1555A→G mutation[J]. J Biol Chem, 2018, 293(9): 3321-3334.
[15] QIAN Y, GUAN M X. Interaction of aminoglycosides with human mitochondrial 12S rRNA carrying the deafness-associated mutation[J]. Antimicrob Agents Chemother, 2009, 53 (11): 4612-4618.
[16] WU L, LI R, CHEN J, et al. Analysis of mitochondrial A1555G mutation in infants with hearing impairment[J]. Exp Ther Med, 2018, 15(6): 5307-5313.
[17] 吴皓, 杨涛. 我国聋病基因的基础研究与临床应用[J]. 中华耳鼻咽喉头颈外科杂志, 2013, 48(12): 969-972.
[18] 何凌. 非综合征型遗传性耳聋基因诊断研究进展[J]. 中国产前诊断杂志(电子版), 2017, 9(4): 47-54.
[19] SCRIMSHAW B J, FAED J M, TATE W P, et al. Rapid identification of an A1555G mutation in human mitochondrial DNA implicated in aminoglycoside-induced ototoxicity[J]. J Hum Genet, 1999, 44(6): 388-390.
[20] RODRÍGUEZ-BALLESTEROS M, OLARTE M, AGUIRRE L A, et al. Molecular and clinical characterisation of three Spanish families with maternally inherited non-syndromic hearing loss caused by the 1494C->T mutation in the mitochondrial 12S rRNA gene[J]. J Med Genet, 2006, 43(11): e54.
[21] BRUIJNS B B, TIGGELAAR R M, GARDENIERS H. The extraction and recovery efficiency of pure DNA for different types of swabs[J]. J Forensic Sci, 2018, 63(5): 1492-1499.
[22] KATILIUS E, CARMEL A B, KOSS H, et al. Sperm cell purification from mock forensic swabs using SOMAmer™ affinity reagents[J]. Forensic Sci Int Genet, 2018, 35: 9-13.
[23] FRANKLN T, PERRY L, SHIH W C, et al. Detection of phytocannabinoids from buccal swabs by headspace solid phase microextraction-gas chromatography/mass spectrome-try[J]. Anal Methods, 2018, 10(9): 942-946.
[24] THEDA C, HWANG S H, CZAJKO A, et al. Quantitation of the cellular content of saliva and buccal swab samples[J]. Sci Rep, 2018, 8(1): 6944.