Application of MALDI-TOF-MS in gene testing for non-syndromic hearing loss
10.3760/cma.j.issn.1673-0860.2013.12.004
- VernacularTitle:飞行时间质谱检测技术在非综合征型耳聋基因检测中的应用
- Author:
Yun ZENG
1
;
Dan JIANG
;
Da-Fei FENG
;
Dong-Dong JIN
;
Xiao-Hui WU
;
Yan-Li DING
;
Jing ZOU
Author Information
1. 518083,深圳华大基因临床检验中心
- Keywords:
Hearing loss;
Mutation;
Spectrometry,mass,matrix-assisted laser desorption-ionization;
Oligonucleotide array sequence analysis
- From:
Chinese Journal of Otorhinolaryngology Head and Neck Surgery
2013;48(12):985-990
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the feasibility of Matrix-Assisted Laser Desorption-Ionization Time of Flight Mass Spectrometry(MALDI-TOF-MS),according to the genetic test of non-syndromic hearing loss (NSHL),and check using the direct sequencing.Methods Peripheral blood was collected from 454NSHL patients.DNA samples were extracted and 20 loci of the four common disease-causing genes were analysed by MALDI-TOF-MS,including GJB2 (35delG,167delT,176_191de116,235delC,299_300delAT),GJB3 (538C→T,547G→A),SLC26A4 (281C→T,589G→A,IVS7-2A→G,1174A→T,1226G→A,1229C→T,IVS15 +5G→A,1975G→C,2027T→A,2162C→T,2168A→G),and mitochondrial 12S rRNA (1494C→T,1555A→G).Direct sequencing was also used to analyse the aforementioned 20 loci in order to validate the accuracy of MALDI-TOF-MS.Results Among the 454 patients,166 cases (36.56％) of disease-causing mutations were detected,which included 69 cases (21.15％) of GJB2 gene mutation,four cases (0.88％) of GJB3 gene mutation,64 cases (14.10％) of SLC26A4 gene mutation,and three cases (0.66％) of mitochondrial 12S rRNA gene mutation.Moreover,the results obtained from direct sequencing and MALDI-TOF-MS were consistent,and the results showed that the two methods were consistent.Conclusions The MALDI-TOF-MS detection method was designed based on the hearing lossrelated mutation hotspots seen in the Chinese population,and it has a high detection rate for NSHL related mutations.In comparison to the conventional detection methods,MALDI-TOF-MS has the following advantages:more detection sites,greater coverage,accurate,high throughput and low cost.Therefore,this method is capable of satisfying the needs of clinical detection for hearing impairment and it is suitable for large-scale implementation.
