Application of PCR reverse dot blot in non-syndromic deafness gene detection
10.13201/j.issn.1001-1781.2020.02.013
- Author:
Yalan LIU
1
;
Shushan SANG
1
;
Jie LING
2
;
Chufeng HE
1
;
Lingyun MEI
1
;
Yong FENG
1
Author Information
1. Department of Otolaryngology Head and Neck Surgery，Xiangya Hospital，Central South University，Changsha，410008，China;Xiangya Hospital，Central South University，Province Key Laboratory of Otolaryngology Critical Diseases
2. Institute of Molecular Precision Medicine，Xiangya Hospital，Central South University
- Publication Type:Journal Article
- Keywords:
hereditary deafness;
PCR;
gene mutation;
molecular hybridization
- From:
Journal of Clinical Otorhinolaryngology Head and Neck Surgery
2020;34(2):153-157
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To detect 20 common deafness gene mutations in non- syndromic deafness patients in China using PCR- RDB, and analyze and summarize the mutation data to explore the clinical value of this method.
Method:The PCR- RDB and Sanger sequencing were used to detect 20 common mutations of four deafness genes（GJB2, GJB3, SLC26A4 and mtDNA） in 500 patients with non- syndromic hearing loss . The Sanger sequencing was used to compare the sensitivity, specificity, positive predictive value, negative predictive value, and total coincidence rate of the deafness mutation detected by PCR- RDB.
Result:A total of 500 samples were detected. 147 wild- type samples, 81 homozygous mutant samples, 240 heterozygous mutant samples, 32 composite heterozygous mutant samples were detected using the PCR- RDB within the range of 20 gene mutations, which were identical to the Sanger sequencing results. GJB2 c.235delC and SLC26A4 c.919- 2 A>G are the most common hotspot mutations in this study, followed by mtDNA m. 1555 A>G. Compared with the Sanger sequencing method, the sensitivity, specificity, positive predictive value, negative predictive value, and total coincidence rate of the real- time fluorescence PCR melting curve method were 100%, and the Kappa value was one.
Conclusion:PCR reverse dot-blot hybridization is a simple, rapid, sensitive and specific method for detecting 20 mutations of 4 common deafness genes in Chinese population, it is expected to be used in clinical detection of deafness genes in the future.
