Application of PCR－reverse dot blot hybridization gene membrane chip technology in genetic testing of children with hereditary nonsyndrome deafness
10.3760/cma.j.issn.2095-428X.2018.23.014
- VernacularTitle:PCR-反向斑点杂交膜芯片技术在遗传性非综合征耳聋患儿基因检测中的应用
- Author:
Minnan YE
1
;
Wenrui LI
;
Qi PENG
;
Siping LI
;
Luheng PENG
;
Xiaomei LU
Author Information
1. 523325,东莞市第八人民医院(东莞市儿童医院)临床检验中心
- Keywords:
Hereditary nonsyndromes deafness;
PCR－reverse dot blot hybridization;
Membrane chips;
Gene mutation
- From:
Chinese Journal of Applied Clinical Pediatrics
2018;33(23):1811-1814
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the application value of PCR－reverse dot blot hybridization (PCR－RDB) gene membrane chip technique in genetic diagnosis of hereditary non－syndrome deafness in children.Methods The blood samples(2 mL)were collected from 38 children with congenital deafness,excluding high－risk factors for deaf-ness at Dongguan Rehabilitation School,and then genomic DNA extracted.By using self－designed multiplex－PCR combined with PCR－RDB gene chip technology,20 hot－spot mutations of 4 pathogenic genes of common deafness in Chinese population were detected.Sanger sequencing was used as the gold standard to corroborate the positive samples. Results Among 38 subjects,deafness gene mutations were detected in 16 cases,with a detection rate of 42.11%,and they were all verified by family study.Among 16 cases,6 cases of GJB2 gene mutation(3 cases of homozygote,3 cases of heterozygous),4 cases of SLC26A4 mutation,2 cases of MTRNR (m.1555A＞G)mutation,4 cases of compound muta-tion,but none of GJB3 gene mutations.And their detection rates were 15.79%,10.53%,5.26%,10.53%,and 0,re-spectively.DNA samples from 16 children with deafness gene mutation were corroborated by Sanger sequencing,and the compliance rate was 100%.Conclusions For 20 hot－spot mutations of 4 common deafness pathogenic genes,the matc-hing PCR－RDB gene membroine chip technology was designed and the susceptible gene of congenital deafness children was detected.This technique has some advantages like high detection rate,fast,accurate and economical.It is an ideal method for gene screening on hereditary non－syndrome deafness children and has good clinical application prospects.
