Background. Deafness is a serious medico-social problem, affecting 8-10% of the population, with approximately 1 in 1000 children suffering from hearing loss. WHO data from 2023 states that more than 430 million people around the globe suffer from hearing loss, and this number is increasing. Non-syndromic Sensorineural Hearing Loss (NSSHL) accounts for 60-80% of childhood genetic deafness. Aim. Local validation of MLPA technique application for molecular analysis of Non-Syndromic Sensorineural Hearing Loss in an independent Moldavian patient. Methods. The implementation and practical application of the molecular-genetic diagnostic method of Non-Syndromic Sensorineural Hearing Loss (NSSHL) and Wolfram syndrome type 1 through the MLPA (Multiplex ligation-dependent probe amplifica-tion) technique allows the early identification of deletions and/or complete duplications of gene exons GJB2, GJB3, GJB6, WFS1, and POU3F4 which are the most common causes of hearing loss in the Republic of Moldova. Results. First of all, the results of the molecular-genetic diagnosis of children with Non-Syndromic Sensorineural Hearing Loss (NSSHL) and Wolfram syndrome type 1 are presented, as well as the effectiveness of prenatal diagnosis in families at risk (Patent Registration no. 6116, 04 August 2023, Chisinau, Nicolae Testemitanu State University of Medicine and Pharmacy, Laboratory of genetics, Mother and Child Institute, Laboratory of Human Molecular Genetics). Conclusions. Molecular-genetic testing, especially for the GJB2 gene of the DFNB1 locus, allows for the initial assessment of any new-born with hypoacusis, to better understand both the cause and the significant impact on the development of the disease, especially for access to specific treatment and care aimed at combating specific types of mutations, as well as genetic counselling and its implications for the family at risk. At the same time, early detected hearing loss, genetic counselling and treatment options, facilitate the recovery of hearing and the optimum development of speech skills in the child.