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|Title:||A Pyrosequencing assay for rapid recognition of SNPs in Mycobacterium tuberculosis embB306 region|
|Citation:||Journal of Microbiological Methods, 2005; 62(1):113-120|
|Publisher:||Elsevier Science BV|
|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.|
|Appears in Collections:||Molecular and Biomedical Science publications|
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