Mitochondrial 12s rRNA mutation A1555G: validation of a genotyping method

BACKGROUND: Research established a genetic basis for aminoglycoside-induced hearing loss (HL) through sequence analysis of the mitochondrial genome, which resulted in the characterization of mutations in the human 12S rRNA that are correlated with aminoglycoside induced ototoxicity. The most common predisposing mutation is the mitochondrial m.1555A>G variant. The aim of the present study was to validate a TaqMAMA genotyping PCR assay MT-RNR1-MAMAPCR real time for detection of mitochondrial m.1555A>G variant. METHODS: To validate the accuracy of the allele-specific real-time PCR genotyping method, we performed both PCR-direct sequencing and TaqMAMA real-time PCR using DNA samples from 54 (29 male and 25 female). Patients who were treated with aminoglycosides mean age, 7.91 +/- 7.12 years were enrolled from September 2019 to January 2020. RESULTS : Precision, Variability and Accuracy were tested. A total of 54 subjects (100%) did not carry any of the tested mutations and was apparently homozygous for the wild-type allele. CONCLUSIONS: The system described here allows accurate and appropriate genotyping in a clinical laboratory setting, allowing the correction of the antibiotic therapy based on the genotyping results of each patient. The emergence of adverse effects after aminoglycoside therapy might be sufficient as an indicator for additional assay of mutations.