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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
Statement of
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.
RMID: 0020094321
DOI: 10.1016/j.mimet.2005.02.004
Appears in Collections:Molecular and Biomedical Science publications

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