Journal
JOURNAL OF ANTIMICROBIAL CHEMOTHERAPY
Volume 69, Issue 4, Pages 947-954Publisher
OXFORD UNIV PRESS
DOI: 10.1093/jac/dkt448
Keywords
tuberculosis; antituberculous resistance; molecular diagnosis; microarrays
Funding
- Spain's Fondo de Investigacion Sanitaria [FIS 06/0328]
- Ministerio de Sanidad y Consumo
- Instituto de Salud Carlos III-FEDER
- Spanish Network for Research in Infectious Diseases [REIPI RD06/0008]
- Institut d'Investigacio Biomedica de Bellvitge (IDIBELL)
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Ethambutol resistance has mostly been related to mutations in the embB gene. The objective of the present study was to characterize the embB gene in a collection of ethambutol-resistant and ethambutol-susceptible isolates of Mycobacterium tuberculosis complex (MTBC) from Barcelona, and to develop a DNA microarray for the rapid detection of embB mutations in our area. Fifty-three ethambutol-resistant and 702 ethambutol-susceptible isolates of MTBC were sequenced in internal 9821495 bp fragments of the embB gene. In addition, a low-cost, low-density array was designed to include the embB codons identified as being most frequently mutated in our area (LD-EMB array). The global prevalence of embB mutations found among the ethambutol-resistant isolates was 77.4 (41/53). Substitutions in embB306 were the most common [53.7 (22/41)], followed by substitutions in embB406 [26.8 (11/41)]. The presence of mutations in embB406 was related to higher levels of ethambutol resistance and to multidrug resistance. Among unrelated isolates (from 24-locus MIRU-VNTR genotyping), the percentage of embB-mutated isolates was 72.9 (27/37)u59.3 (16/27) in embB306 and 25.9 (7/27) in embB406. None of the ethambutol-susceptible isolates studied showed a mutation in codon 306 or 406. The LD-EMB array showed 100 sensitivity and specificity in identifying the main embB substitutions in our area. Mutations at codons 306 and 406 of embB have a relevant role in resistance to ethambutol in our area. The LD-EMB array developed in this study would appear to be a good molecular test for rapid detection of ethambutol resistance.
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