alexa Rapid and Accurate Mitochondrial DNA Analysis in Amino Glycoside Sensitive Patients
ISSN: 2161-1009

Biochemistry & Analytical Biochemistry
Open Access

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Research Article

Rapid and Accurate Mitochondrial DNA Analysis in Amino Glycoside Sensitive Patients

Torres-Ruiz NM and Meza G*
Division of Neuroscience, Institute of Cellular Physiology, National Autonomous University of Mexico, Mexico DF 04510, Mexico
Corresponding Author : Graciela Meza
Division of Neuroscience, Institute of Cellular Physiology
National Autonomous University of Mexico
Apartado Postal 70-253, 04510, Mexico
Tel: (52) 56-22-55-85
Fax: (52) 56-22-57-47
E-mail: [email protected]
Received February 08, 2012; Accepted March 21, 2012; Published March 23, 2012
Citation: Torres-Ruiz NM, Meza G (2012) Rapid and Accurate Mitochondrial DNA Analysis in Amino Glycoside Sensitive Patients. Biochem & Anal Biochem S3:002. doi:10.4172/2161-1009.S3-002
Copyright: © 2012 Torres-Ruiz NM, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
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Abstract

The use of Restriction Fragment Length Polymorphism to assess the position of mutation in nuclear or mitochondrial DNA is very common but the effectiveness of this methodology is limited and expensive since it utilizes many reagents that are not easy to acquire. We propose the use of a Denaturing Gradients Gel Electrophoresis to analyze the presence of mutation T1189C in the 12S rRNA region of human mitochondrial DNA, formerly detected by nucleotide sequence analysis, in order to develop an optimized method that would allow the simultaneous detection of this mutation in several patient mitochondrial DNA and further finding the relationship of its presence with sudden deafness produced by hypersensitivity to aminoglycoside antibiotic treatment. The technique was improved optimizing the denaturing gradient gel electrophoresis parameters, such as optimum temperature, voltage, concentration of denaturing agents and time for carrying out the method, which allowed us to precisely distinguish whether there has been a change in the sequence of a given sample, analyzing a wild type mitochondrial 12S rRNA against the patient sample simultaneously, by simply observing the differences in running time of a band in which mutation is present, and corroborated by sequence analysis. The application of the technique to samples of various patients at the same time would be very valuable to assess the presence of the mutation prior to treatment of a given infection is started and to recommend the use of an alternative therapeutic agent innocuous for the inner ear.

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