Multiplex allele-specific PCR assays for the identification of mitochondrial 12S rRNA mutations
10.3760/cma.j.issn.1009-9158.2011.07.012
- VernacularTitle:多重等位基因PCR检测线粒体12S rRNA基因突变
- Author:
Jing ZHENG
;
Aifen YANG
;
Ting ZHANG
;
Qiongmin ZHANG
;
Shasha GONG
;
Guanghua PENG
;
Yi ZHU
;
Minxin GUAN
- Publication Type:Journal Article
- Keywords:
Deafness,mitochondria;
Aminoglycosides;
RNA,ribosomal;
Mutation;
Polymerase chain reaction
- From:
Chinese Journal of Laboratory Medicine
2011;34(7):628-632
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the clinical application of multiplex allele-specific PCR assays for simultaneous detection of the mitochondrial 12S rRNA A1555G and C1494T mutations associated with aminoglycoside-induced hearing impairment.Methods Three standard plasmids of different genotypes (wild-type, A1555G mutant and C1494T mutant) were constructed for templates and allele-specific primers aiming directly at wild-type and mutant of mitochondrial DNA nt1555 and nt1494 were designed for developing a multiplex allele-specific PCR technique to detect the A1555G and C1494T mutations.Then the method was applied to clinical screening of 138 non-syndromic hearing loss subjects and confirmed by DNA sequencing.Results Multiplex allele-specific PCR was successfully applied to the detection of A1555G and C1494T mutations in a cohort of 138 Han Chinese genetically unrelated hearing-loss subjects.Finally, 11(7.97%) unrelated affected subjects harbored the A1555G and C1494T mutations in the 12S rRNA gene(10 cases for A1555G and 1 cases for C1494T), which was well consistent with results of DNA sequencing [7.97%(11/138), Kappa=1.000, P＜0.01].Conclusion This study indicates that the multiplex allele-specific PCR assay is useful, convenient and reliable in the detection of the A1555G and C1494T mutations, which could identify the subjects at risk and effectively prevent of aminoglycoside-induced hearing loss.
