TY - JOUR
T1 - Pyrosequencing for rapid detection of Mycobacterium tuberculosis second-line drugs and ethambutol resistance
AU - Lacoma, Alicia
AU - Molina-Moya, Barbara
AU - Prat, Cristina
AU - Pimkina, Edita
AU - Diaz, Jessica
AU - Dudnyk, Andriy
AU - García-Sierra, Nerea
AU - Haba, Lucía
AU - Maldonado, Jose
AU - Samper, Sofia
AU - Ruiz-Manzano, Juan
AU - Ausina, Vicente
AU - Dominguez, Jose
N1 - Funding Information:
This work was supported by grants from Fondo de Investigaciones Sanitarias ( FIS07/0551 ) (Ministerio de Sanidad y Consumo, Madrid, Spain) and by the Spanish Society of Pneumology and Thoracic Surgery. J. Domínguez is funded by the “Miguel Servet” program of the Instituto de Salud Carlos III (Spain). A. Dudnyk is a recipient of a Short-Term Fellowship grant from European Respiratory Society ( STRTF – 51-2012 ).
Publisher Copyright:
© 2015 Elsevier Inc..
PY - 2015/11
Y1 - 2015/11
N2 - 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 MTBDR. sl (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.
AB - 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 MTBDR. sl (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.
KW - Ethambutol
KW - Extensively drug-resistant tuberculosis
KW - Fluoroquinolones
KW - Injectable drugs
KW - Molecular diagnostic testing
KW - Tuberculosis
UR - http://www.scopus.com/inward/record.url?scp=84943583948&partnerID=8YFLogxK
U2 - 10.1016/j.diagmicrobio.2015.07.004
DO - 10.1016/j.diagmicrobio.2015.07.004
M3 - Article
C2 - 26256417
AN - SCOPUS:84943583948
SN - 0732-8893
VL - 83
SP - 263
EP - 269
JO - Diagnostic Microbiology and Infectious Disease
JF - Diagnostic Microbiology and Infectious Disease
IS - 3
ER -