Rapid identification and susceptibility testing of Mycobacterium tuberculosis from MGIT cultures with luciferase reporter mycobacteriophages

Niaz Banaiee, Miriam Bobadilla-del-Valle, Paul F. Riska, Svetoslav Bardarov, Peter M. Small, Alfredo Ponce-de-Leon, William R. Jacobs, Graham F. Hatfull, Jose Sifuentes-Osornio

Research output: Contribution to journalArticle

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Abstract

In a prospective study conducted in a diagnostic laboratory in Mexico City, luciferase reporter mycobacteriophages (LRPs) were evaluated for their utility and performance in identification and antibiotic-susceptibility testing of Mycobacterium tuberculosis complex (MTC) isolates from MGIT-960 cultures. Eighty-four consecutive MGIT cultures recovered from 54 patients were included in this study. The LRPs confirmed mycobacterial growth in 79 (94%) of 84 MGIT cultures. Failure to confirm growth was due to low inoculum (n = 1) or growth with non-tuberculous mycobacteria (n = 4). The median time to confirmation of MGIT cultures was 1 day (range 1-55). Confirmed cultures were identified with p-nitro-α-acetylamino-β-hydroxypropiophenone (NAP), a selective inhibitor of MTC species, and results obtained with LRPs were compared with those obtained by BACTEC-460. The sensitivity and specificity of the LRP NAP test were respectively 97 and 100%, and the median turnaround time for identification was 3 days with both methods. The accuracy and speed of the LRPs for susceptibility testing with rifampicin, streptomycin, isoniazid and ethambutol were compared with BACTEC-460 and discrepant results were tested by the conventional agar proportion method. In total, 72 MTC cultures were tested. The overall agreement between the LRPs and BACTEC-460 was 98.6%. Four isolates (5.6%) were falsely identified as ethambutol-resistant. The median turnaround time for susceptibility testing was 3 days (range 3-57) with the LRPs and 9 days (range 7-29) with BACTEC-460. LRPs offer an accurate and rapid approach for identification and susceptibility testing of M. tuberculosis from MGIT-960 cultures.

Original languageEnglish (US)
Pages (from-to)557-561
Number of pages5
JournalJournal of Medical Microbiology
Volume52
Issue number7
DOIs
StatePublished - Jul 1 2003
Externally publishedYes

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Mycobacteriophages
Luciferases
Mycobacterium tuberculosis
Ethambutol
Hydroxypropiophenone
Growth
Isoniazid
Streptomycin
Rifampin
Mycobacterium
Mexico
Agar
Prospective Studies
Anti-Bacterial Agents
Sensitivity and Specificity

ASJC Scopus subject areas

  • Microbiology (medical)
  • Microbiology

Cite this

Rapid identification and susceptibility testing of Mycobacterium tuberculosis from MGIT cultures with luciferase reporter mycobacteriophages. / Banaiee, Niaz; Bobadilla-del-Valle, Miriam; Riska, Paul F.; Bardarov, Svetoslav; Small, Peter M.; Ponce-de-Leon, Alfredo; Jacobs, William R.; Hatfull, Graham F.; Sifuentes-Osornio, Jose.

In: Journal of Medical Microbiology, Vol. 52, No. 7, 01.07.2003, p. 557-561.

Research output: Contribution to journalArticle

Banaiee, N, Bobadilla-del-Valle, M, Riska, PF, Bardarov, S, Small, PM, Ponce-de-Leon, A, Jacobs, WR, Hatfull, GF & Sifuentes-Osornio, J 2003, 'Rapid identification and susceptibility testing of Mycobacterium tuberculosis from MGIT cultures with luciferase reporter mycobacteriophages', Journal of Medical Microbiology, vol. 52, no. 7, pp. 557-561. https://doi.org/10.1099/jmm.0.05149-0
Banaiee, Niaz ; Bobadilla-del-Valle, Miriam ; Riska, Paul F. ; Bardarov, Svetoslav ; Small, Peter M. ; Ponce-de-Leon, Alfredo ; Jacobs, William R. ; Hatfull, Graham F. ; Sifuentes-Osornio, Jose. / Rapid identification and susceptibility testing of Mycobacterium tuberculosis from MGIT cultures with luciferase reporter mycobacteriophages. In: Journal of Medical Microbiology. 2003 ; Vol. 52, No. 7. pp. 557-561.
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AU - Bobadilla-del-Valle, Miriam

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AU - Small, Peter M.

AU - Ponce-de-Leon, Alfredo

AU - Jacobs, William R.

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AU - Sifuentes-Osornio, Jose

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N2 - In a prospective study conducted in a diagnostic laboratory in Mexico City, luciferase reporter mycobacteriophages (LRPs) were evaluated for their utility and performance in identification and antibiotic-susceptibility testing of Mycobacterium tuberculosis complex (MTC) isolates from MGIT-960 cultures. Eighty-four consecutive MGIT cultures recovered from 54 patients were included in this study. The LRPs confirmed mycobacterial growth in 79 (94%) of 84 MGIT cultures. Failure to confirm growth was due to low inoculum (n = 1) or growth with non-tuberculous mycobacteria (n = 4). The median time to confirmation of MGIT cultures was 1 day (range 1-55). Confirmed cultures were identified with p-nitro-α-acetylamino-β-hydroxypropiophenone (NAP), a selective inhibitor of MTC species, and results obtained with LRPs were compared with those obtained by BACTEC-460. The sensitivity and specificity of the LRP NAP test were respectively 97 and 100%, and the median turnaround time for identification was 3 days with both methods. The accuracy and speed of the LRPs for susceptibility testing with rifampicin, streptomycin, isoniazid and ethambutol were compared with BACTEC-460 and discrepant results were tested by the conventional agar proportion method. In total, 72 MTC cultures were tested. The overall agreement between the LRPs and BACTEC-460 was 98.6%. Four isolates (5.6%) were falsely identified as ethambutol-resistant. The median turnaround time for susceptibility testing was 3 days (range 3-57) with the LRPs and 9 days (range 7-29) with BACTEC-460. LRPs offer an accurate and rapid approach for identification and susceptibility testing of M. tuberculosis from MGIT-960 cultures.

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