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https://hdl.handle.net/2440/55380
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Type: | Journal article |
Title: | A Pyrosequencing assay for rapid recognition of SNPs in Mycobacterium tuberculosis embB306 region |
Author: | Isola, D. Pardini, M. Varaine, F. Niemann, S. Rusch-Gerdes, S. Fattorini, L. Orefici, G. Meacci, F. Trappetti, C. Oggioni, M. Orru, G. |
Citation: | Journal of Microbiological Methods, 2005; 62(1):113-120 |
Publisher: | Elsevier Science BV |
Issue Date: | 2005 |
ISSN: | 0167-7012 1872-8359 |
Statement of Responsibility: | Daniela Isolaa, Manuela Pardinib, Francis Varainec, Stefan Niemanne, Sabine Rüsch-Gerdese, Lanfranco Fattorinib, Graziella Oreficib, Francesca Meaccid, Claudia Trappettid, Marco Rinaldo Oggionid, the LONG-DRUG study group, Germano Orrù |
Abstract: | Ethambutol (EMB) is in use worldwide as a first-line anti-tuberculosis drug and substitutions in codon 306 of the embB gene are the most common mutations found in EMB resistant Mycobacterium tuberculosis (MTB) strains. Pyrosequencing is a real time sequencing method able to rapidly detect mutations in a large number of samples. Using this technique we analyzed, in parallel with conventional sequencing, a 24 bp region of the embB gene of 28 MTB clinical isolates. Pyrosequencing efficiently identified all embB306 mutations, detecting three different single-base substitutions leading to 2 amino acid changes (Met to Val or Ile). Mutated embB alleles were detected in 2 multidrug-resistant (MDR) EMB-susceptible strains. Overall, our results demonstrated that the Pyrosequencing method efficiently recognizes mutations in embB in a very short time and represents a valid molecular method to detect mutations in the MTB embB306 region. |
Keywords: | Ethambutol embB Mycobacterium tuberculosis Pyrosequencing method |
Description: | Copyright © 2005 Elsevier B.V. All rights reserved. |
DOI: | 10.1016/j.mimet.2005.02.004 |
Published version: | http://dx.doi.org/10.1016/j.mimet.2005.02.004 |
Appears in Collections: | Aurora harvest 5 Molecular and Biomedical Science publications |
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