title: Molecular Analysis of the embCAB Locus and embR Gene Involved in Ethambutol Resistance in Clinical Isolates of Mycobacterium tuberculosis in France.
abstract: Modification of codon 306 in embB is regarded as the main mechanism leading to ethambutol (ETB) resistance in clinical isolates of Mycobacterium tuberculosis. However, numerous mutations elsewhere in the embCAB locus and in embR, a putative transcriptional activator of this locus, have been reported to be involved in ETB resistance. Here, we investigated the diversity of nucleotide variations observed in embCAB and embR in M. tuberculosis complex isolates from France. These regions were sequenced in 71 ETB-resistant (ETB-R) and 60 ETB-susceptible (ETB-S) clinical isolates of known phylogenetic lineages. The 131 isolates had 12 mutations corresponding to phylogenetic markers. Among the 60 ETB-S isolates, only 3 (5%) had nonsynonymous mutations that were not phylogenetic markers. Among the 71 ETB-R isolates, 98% had mutations in embCAB that likely contribute to ETB resistance: 70% had mutations located in embB codon 306, 406, or 497; 13% had mutations located outside these three positions between codons 296 and 426; and 15% had mutations corresponding to mutations in the embC-embA intergenic region. We found a strong association between resistance to ETB and the presence of mutations in embB and the embC-embA intergenic region (P < 0.001). In contrast, the mutations detected in embC and embA were not involved in ETB resistance, and no mutation was detected in embR. These results strongly suggest that the sensitivity of diagnostic assays for detecting ETB resistance based on testing of embB codon 306 can be increased by testing of the embB region between codons 296 and 497 and by including the embC-embA intergenic region between positions -8 and -21.
authors.affiliation: AP-HP, Hôpital Pitié-Salpêtrière, Centre National de Référence des Mycobactéries et de la Résistance des Mycobactéries aux Antituberculeux, Paris, France Sorbonne Universités, UPMC Université Paris 06, UMRS1135, Centre d'Immunologie et des Maladies Infectieuses, CIMI-Paris, Team 13 (Bacteriology), Paris, France INSERM, U1135, Centre d'Immunologie et des Maladies Infectieuses, CIMI-Paris, Team 13 (Bacteriology), Paris, France florence.brossier@psl.aphp.fr.
authors.forename: Florence
authors.initials: F
authors.lastname: Brossier
authors.affiliation: AP-HP, Hôpital Pitié-Salpêtrière, Centre National de Référence des Mycobactéries et de la Résistance des Mycobactéries aux Antituberculeux, Paris, France Sorbonne Universités, UPMC Université Paris 06, UMRS1135, Centre d'Immunologie et des Maladies Infectieuses, CIMI-Paris, Team 13 (Bacteriology), Paris, France INSERM, U1135, Centre d'Immunologie et des Maladies Infectieuses, CIMI-Paris, Team 13 (Bacteriology), Paris, France.
authors.forename: Wladimir
authors.initials: W
authors.lastname: Sougakoff
authors.affiliation: AP-HP, Hôpital Pitié-Salpêtrière, Centre National de Référence des Mycobactéries et de la Résistance des Mycobactéries aux Antituberculeux, Paris, France Sorbonne Universités, UPMC Université Paris 06, UMRS1135, Centre d'Immunologie et des Maladies Infectieuses, CIMI-Paris, Team 13 (Bacteriology), Paris, France INSERM, U1135, Centre d'Immunologie et des Maladies Infectieuses, CIMI-Paris, Team 13 (Bacteriology), Paris, France.
authors.forename: Christine
authors.initials: C
authors.lastname: Bernard
authors.affiliation: Institut de Génétique et Microbiologie, UMR8621 CNRS-Université Paris-Sud 11, UniverSud, Infection Genetics Emerging Pathogens Evolution (IGEPE) Team, Orsay, France.
authors.forename: Matthieu
authors.initials: M
authors.lastname: Petrou
authors.affiliation: Sorbonne Universités, UPMC Université Paris 06, UMRS1135, Centre d'Immunologie et des Maladies Infectieuses, CIMI-Paris, Team 13 (Bacteriology), Paris, France INSERM, U1135, Centre d'Immunologie et des Maladies Infectieuses, CIMI-Paris, Team 13 (Bacteriology), Paris, France.
authors.forename: Karine
authors.initials: K
authors.lastname: Adeyema
authors.affiliation: Sorbonne Universités, UPMC Université Paris 06, UMRS1135, Centre d'Immunologie et des Maladies Infectieuses, CIMI-Paris, Team 13 (Bacteriology), Paris, France INSERM, U1135, Centre d'Immunologie et des Maladies Infectieuses, CIMI-Paris, Team 13 (Bacteriology), Paris, France.
authors.forename: Anne
authors.initials: A
authors.lastname: Pham
authors.affiliation: Sorbonne Universités, UPMC Université Paris 06, UMRS1135, Centre d'Immunologie et des Maladies Infectieuses, CIMI-Paris, Team 13 (Bacteriology), Paris, France INSERM, U1135, Centre d'Immunologie et des Maladies Infectieuses, CIMI-Paris, Team 13 (Bacteriology), Paris, France.
authors.forename: Diane
authors.initials: D
authors.lastname: Amy de la Breteque
authors.affiliation: Sorbonne Universités, UPMC Université Paris 06, UMRS1135, Centre d'Immunologie et des Maladies Infectieuses, CIMI-Paris, Team 13 (Bacteriology), Paris, France INSERM, U1135, Centre d'Immunologie et des Maladies Infectieuses, CIMI-Paris, Team 13 (Bacteriology), Paris, France.
authors.forename: Marine
authors.initials: M
authors.lastname: Vallet
authors.affiliation: AP-HP, Hôpital Pitié-Salpêtrière, Centre National de Référence des Mycobactéries et de la Résistance des Mycobactéries aux Antituberculeux, Paris, France Sorbonne Universités, UPMC Université Paris 06, UMRS1135, Centre d'Immunologie et des Maladies Infectieuses, CIMI-Paris, Team 13 (Bacteriology), Paris, France INSERM, U1135, Centre d'Immunologie et des Maladies Infectieuses, CIMI-Paris, Team 13 (Bacteriology), Paris, France.
authors.forename: Vincent
authors.initials: V
authors.lastname: Jarlier
authors.affiliation: Institut de Génétique et Microbiologie, UMR8621 CNRS-Université Paris-Sud 11, UniverSud, Infection Genetics Emerging Pathogens Evolution (IGEPE) Team, Orsay, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Sola
authors.affiliation: AP-HP, Hôpital Pitié-Salpêtrière, Centre National de Référence des Mycobactéries et de la Résistance des Mycobactéries aux Antituberculeux, Paris, France Sorbonne Universités, UPMC Université Paris 06, UMRS1135, Centre d'Immunologie et des Maladies Infectieuses, CIMI-Paris, Team 13 (Bacteriology), Paris, France INSERM, U1135, Centre d'Immunologie et des Maladies Infectieuses, CIMI-Paris, Team 13 (Bacteriology), Paris, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Veziris
mesh_terms: D000995:Antitubercular Agents / Q000494:pharmacology*; D003062:Codon / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D021901:DNA, Intergenic / Q000235:genetics; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D004977:Ethambutol / Q000494:pharmacology*; D005798:Genes, Bacterial / Q000235:genetics*; D014644:Genetic Variation / Q000235:genetics; D006801:Humans; D008826:Microbial Sensitivity Tests / Q000379:methods; D009154:Mutation / Q000235:genetics; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000235:genetics*; D010802:Phylogeny; D015534:Trans-Activators / Q000235:genetics
chemical_list: D000995:Antitubercular Agents; D003062:Codon; D004269:DNA, Bacterial; D021901:DNA, Intergenic; D015534:Trans-Activators; D004977:Ethambutol