| Literature DB >> 15823399 |
Daniela Isola1, Manuela Pardini, Francis Varaine, Stefan Niemann, Sabine Rüsch-Gerdes, Lanfranco Fattorini, Graziella Orefici, Francesca Meacci, Claudia Trappetti, Marco Rinaldo Oggioni, 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.Entities:
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Year: 2005 PMID： 15823399 DOI： 10.1016/j.mimet.2005.02.004
Source DB: PubMed Journal: J Microbiol Methods ISSN： 0167-7012 Impact factor: 2.363