title: Pyrosequencing for rapid detection of Mycobacterium tuberculosis second-line drugs and ethambutol resistance.
abstract: The aim of this work was to study the diagnostic accuracy of pyrosequencing to detect resistance to fluoroquinolones, kanamycin, amikacin, capreomycin, and ethambutol (EMB) in Mycobacterium tuberculosis clinical strains. One hundred four clinical isolates previously characterized by BACTEC 460TB/MGIT 960 were included. Specific mutations were targeted in gyrA, rrs, eis promoter, and embB. When there was a discordant result between BACTEC and pyrosequencing, Genotype MTBDRsl (Hain Lifescience, Nehren, Germany) was performed. Sensitivity and specificity of pyrosequencing were 70.6% and 100%, respectively, for fluoroquinolones; 93.3% and 81.7%, respectively, for kanamycin; 94.1% and 95.9%, respectively, for amikacin; 90.0% and 100%, respectively, for capreomycin; and 64.8% and 87.8%, respectively, for EMB. This study shows that pyrosequencing may be a useful tool for making early decisions regarding second-line drugs and EMB resistance. However, for a correct management of patients with suspected extensively drug-resistant tuberculosis, susceptibility results obtained by molecular methods should be confirmed by a phenotypic method.
authors.affiliation: Servei de Microbiologia, Hospital Universitari Germans Trias i Pujol, Institut d'Investigació Germans Trias i Pujol, Universitat Autònoma de Barcelona, Carretera del Canyet s/n, 08916, Badalona, Spain; CIBER Enfermedades Respiratorias (CIBERES), Instituto de Salud Carlos III, Spain.
authors.forename: Alicia
authors.initials: A
authors.lastname: Lacoma
authors.affiliation: Servei de Microbiologia, Hospital Universitari Germans Trias i Pujol, Institut d'Investigació Germans Trias i Pujol, Universitat Autònoma de Barcelona, Carretera del Canyet s/n, 08916, Badalona, Spain; CIBER Enfermedades Respiratorias (CIBERES), Instituto de Salud Carlos III, Spain.
authors.forename: Barbara
authors.initials: B
authors.lastname: Molina-Moya
authors.affiliation: Servei de Microbiologia, Hospital Universitari Germans Trias i Pujol, Institut d'Investigació Germans Trias i Pujol, Universitat Autònoma de Barcelona, Carretera del Canyet s/n, 08916, Badalona, Spain; CIBER Enfermedades Respiratorias (CIBERES), Instituto de Salud Carlos III, Spain.
authors.forename: Cristina
authors.initials: C
authors.lastname: Prat
authors.affiliation: Infectious Diseases and Tuberculosis Hospital, Affiliate of Vilnius University Hospital Santariskiu Klinikos, Vilnius, Lithuania.
authors.forename: Edita
authors.initials: E
authors.lastname: Pimkina
authors.affiliation: Servei de Microbiologia, Hospital Universitari Germans Trias i Pujol, Institut d'Investigació Germans Trias i Pujol, Universitat Autònoma de Barcelona, Carretera del Canyet s/n, 08916, Badalona, Spain; CIBER Enfermedades Respiratorias (CIBERES), Instituto de Salud Carlos III, Spain.
authors.forename: Jessica
authors.initials: J
authors.lastname: Diaz
authors.affiliation: Department of Tuberculosis, Clinical Immunology and Allergology, Vinnitsa National Pirogov Memorial Medical University, Vinnitsa, Ukraine.
authors.forename: Andriy
authors.initials: A
authors.lastname: Dudnyk
authors.affiliation: Servei de Microbiologia, Hospital Universitari Germans Trias i Pujol, Institut d'Investigació Germans Trias i Pujol, Universitat Autònoma de Barcelona, Carretera del Canyet s/n, 08916, Badalona, Spain.
authors.forename: Nerea
authors.initials: N
authors.lastname: García-Sierra
authors.affiliation: Servei de Microbiologia, Hospital Universitari Germans Trias i Pujol, Institut d'Investigació Germans Trias i Pujol, Universitat Autònoma de Barcelona, Carretera del Canyet s/n, 08916, Badalona, Spain.
authors.forename: Lucía
authors.initials: L
authors.lastname: Haba
authors.affiliation: Serveis Clínics, Barcelona, Spain.
authors.forename: Jose
authors.initials: J
authors.lastname: Maldonado
authors.affiliation: CIBER Enfermedades Respiratorias (CIBERES), Instituto de Salud Carlos III, Spain; Instituto Aragonés de Ciencias de la Salud, Hospital Universitario Miguel Servet, Zaragoza, Spain.
authors.forename: Sofia
authors.initials: S
authors.lastname: Samper
authors.affiliation: Servei de Pneumologia, Hospital Universitari Germans Trias i Pujol, Institut d'Investigació Germans Trias i Pujol, Universitat Autònoma de Barcelona, Carretera del Canyet s/n, 08916, Badalona, Spain; CIBER Enfermedades Respiratorias (CIBERES), Instituto de Salud Carlos III, Spain.
authors.forename: Juan
authors.initials: J
authors.lastname: Ruiz-Manzano
authors.affiliation: Servei de Microbiologia, Hospital Universitari Germans Trias i Pujol, Institut d'Investigació Germans Trias i Pujol, Universitat Autònoma de Barcelona, Carretera del Canyet s/n, 08916, Badalona, Spain; CIBER Enfermedades Respiratorias (CIBERES), Instituto de Salud Carlos III, Spain.
authors.forename: Vicente
authors.initials: V
authors.lastname: Ausina
authors.affiliation: Servei de Microbiologia, Hospital Universitari Germans Trias i Pujol, Institut d'Investigació Germans Trias i Pujol, Universitat Autònoma de Barcelona, Carretera del Canyet s/n, 08916, Badalona, Spain; CIBER Enfermedades Respiratorias (CIBERES), Instituto de Salud Carlos III, Spain. Electronic address: jadomb@gmail.com.
authors.forename: Jose
authors.initials: J
authors.lastname: Dominguez
mesh_terms: D000995:Antitubercular Agents / Q000494:pharmacology*; D024881:Drug Resistance, Bacterial; D060005:Genotyping Techniques / Q000379:methods*; D006801:Humans; D008826:Microbial Sensitivity Tests / Q000379:methods*; D009169:Mycobacterium tuberculosis / Q000187:drug effects / Q000235:genetics*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000379:methods*; D013997:Time Factors
chemical_list: D000995:Antitubercular Agents
keywords: Ethambutol; Extensively drug-resistant tuberculosis; Fluoroquinolones; Injectable drugs; Molecular diagnostic testing; Tuberculosis