Luciferase reporter mycobacteriophages for detection, identification, and antibiotic susceptibility testing of mycobacterium tuberculosis in Mexico

N. Banaiee, M. Bobadilla-del-Valle, Jr Bardarov S., P. F. Riska, P. M. Small, A. Ponce-de-Leon, Jr Jacobs W.R., G. F. Hatfull, J. Sifuentes-Osornio

Research output: Contribution to journalArticle

57 Citations (Scopus)

Abstract

The utility of luciferase reporter mycobacteriophages (LRPs) for detection, identification, and antibiotic susceptibility testing of Mycobacterium tuberculosis was prospectively evaluated in a clinical microbiology laboratory in Mexico City, Mexico. Five hundred twenty-three consecutive sputum samples submitted to the laboratory during a 5-month period were included in this study. These specimens were cultivated in Middle-brook 7H9 (MADC), MGIT, and Löwenstein-Jensen (LJ) media. Of the 71 mycobacterial isolates recovered with any of the three media, 76% were detected with the LRPs, 97% were detected with the MGIT 960 method, and 90% were detected with LJ medium. When contaminated specimens were excluded from the analysis, the LRPs detected 92% (54 of 59) of the cultures. The median time to detection of bacteria was 7 days with both the LRPs and the MGIT 960 method. LRP detection of growth in the presence of p-nitro-α-acetylamino-β-hydroxypropiophenone (NAP) was used for selective identification of M. tuberculosis complex (MTC) and compared to identification with BACTEC 460. Using the LRP NAP test, 47 (94%) out of 50 isolates were correctly identified as tuberculosis complex. The accuracy and speed of LRP antibiotic susceptibility testing with rifampin, streptomycin, isoniazid, and ethambutol were compared to those of the BACTEC 460 method, and discrepant results were checked by the conventional proportion method. In total, 50 MTC isolates were tested. The overall agreement between the LRP and BACTEC 460 results was 98.5%. The median LRP-based susceptibility turnaround time was 2 days (range, 2 to 4 days) compared to 10.5 days (range, 7 to 16 days) by the BACTEC 460 method. Phage resistance was not detected in any of the 243 MTC isolates tested. Mycobacteriophage-based approaches to tuberculosis diagnostics can be implemented in clinical laboratories with sensitivity, specificity, and rapidity that compare favorably with those of the MGIT 960 and BACTEC 460 methods. The phages currently provide the fastest phenotypic assay for susceptibility testing.

Original languageEnglish (US)
Pages (from-to)3883-3888
Number of pages6
JournalJournal of Clinical Microbiology
Volume39
Issue number11
DOIs
StatePublished - 2001
Externally publishedYes

Fingerprint

Mycobacteriophages
Mexico
Luciferases
Mycobacterium tuberculosis
Anti-Bacterial Agents
Bacteriophages
Hydroxypropiophenone
Tuberculosis
Ethambutol
Isoniazid
Streptomycin
Rifampin
Microbiology
Sputum

ASJC Scopus subject areas

  • Microbiology (medical)
  • Microbiology

Cite this

Luciferase reporter mycobacteriophages for detection, identification, and antibiotic susceptibility testing of mycobacterium tuberculosis in Mexico. / Banaiee, N.; Bobadilla-del-Valle, M.; Bardarov S., Jr; Riska, P. F.; Small, P. M.; Ponce-de-Leon, A.; Jacobs W.R., Jr; Hatfull, G. F.; Sifuentes-Osornio, J.

In: Journal of Clinical Microbiology, Vol. 39, No. 11, 2001, p. 3883-3888.

Research output: Contribution to journalArticle

Banaiee, N. ; Bobadilla-del-Valle, M. ; Bardarov S., Jr ; Riska, P. F. ; Small, P. M. ; Ponce-de-Leon, A. ; Jacobs W.R., Jr ; Hatfull, G. F. ; Sifuentes-Osornio, J. / Luciferase reporter mycobacteriophages for detection, identification, and antibiotic susceptibility testing of mycobacterium tuberculosis in Mexico. In: Journal of Clinical Microbiology. 2001 ; Vol. 39, No. 11. pp. 3883-3888.
@article{a08a5394c2244db8a210839f04180ad5,
title = "Luciferase reporter mycobacteriophages for detection, identification, and antibiotic susceptibility testing of mycobacterium tuberculosis in Mexico",
abstract = "The utility of luciferase reporter mycobacteriophages (LRPs) for detection, identification, and antibiotic susceptibility testing of Mycobacterium tuberculosis was prospectively evaluated in a clinical microbiology laboratory in Mexico City, Mexico. Five hundred twenty-three consecutive sputum samples submitted to the laboratory during a 5-month period were included in this study. These specimens were cultivated in Middle-brook 7H9 (MADC), MGIT, and L{\"o}wenstein-Jensen (LJ) media. Of the 71 mycobacterial isolates recovered with any of the three media, 76{\%} were detected with the LRPs, 97{\%} were detected with the MGIT 960 method, and 90{\%} were detected with LJ medium. When contaminated specimens were excluded from the analysis, the LRPs detected 92{\%} (54 of 59) of the cultures. The median time to detection of bacteria was 7 days with both the LRPs and the MGIT 960 method. LRP detection of growth in the presence of p-nitro-α-acetylamino-β-hydroxypropiophenone (NAP) was used for selective identification of M. tuberculosis complex (MTC) and compared to identification with BACTEC 460. Using the LRP NAP test, 47 (94{\%}) out of 50 isolates were correctly identified as tuberculosis complex. The accuracy and speed of LRP antibiotic susceptibility testing with rifampin, streptomycin, isoniazid, and ethambutol were compared to those of the BACTEC 460 method, and discrepant results were checked by the conventional proportion method. In total, 50 MTC isolates were tested. The overall agreement between the LRP and BACTEC 460 results was 98.5{\%}. The median LRP-based susceptibility turnaround time was 2 days (range, 2 to 4 days) compared to 10.5 days (range, 7 to 16 days) by the BACTEC 460 method. Phage resistance was not detected in any of the 243 MTC isolates tested. Mycobacteriophage-based approaches to tuberculosis diagnostics can be implemented in clinical laboratories with sensitivity, specificity, and rapidity that compare favorably with those of the MGIT 960 and BACTEC 460 methods. The phages currently provide the fastest phenotypic assay for susceptibility testing.",
author = "N. Banaiee and M. Bobadilla-del-Valle and {Bardarov S.}, Jr and Riska, {P. F.} and Small, {P. M.} and A. Ponce-de-Leon and {Jacobs W.R.}, Jr and Hatfull, {G. F.} and J. Sifuentes-Osornio",
year = "2001",
doi = "10.1128/JCM.39.11.3883-3888.2001",
language = "English (US)",
volume = "39",
pages = "3883--3888",
journal = "Journal of Clinical Microbiology",
issn = "0095-1137",
publisher = "American Society for Microbiology",
number = "11",

}

TY - JOUR

T1 - Luciferase reporter mycobacteriophages for detection, identification, and antibiotic susceptibility testing of mycobacterium tuberculosis in Mexico

AU - Banaiee, N.

AU - Bobadilla-del-Valle, M.

AU - Bardarov S., Jr

AU - Riska, P. F.

AU - Small, P. M.

AU - Ponce-de-Leon, A.

AU - Jacobs W.R., Jr

AU - Hatfull, G. F.

AU - Sifuentes-Osornio, J.

PY - 2001

Y1 - 2001

N2 - The utility of luciferase reporter mycobacteriophages (LRPs) for detection, identification, and antibiotic susceptibility testing of Mycobacterium tuberculosis was prospectively evaluated in a clinical microbiology laboratory in Mexico City, Mexico. Five hundred twenty-three consecutive sputum samples submitted to the laboratory during a 5-month period were included in this study. These specimens were cultivated in Middle-brook 7H9 (MADC), MGIT, and Löwenstein-Jensen (LJ) media. Of the 71 mycobacterial isolates recovered with any of the three media, 76% were detected with the LRPs, 97% were detected with the MGIT 960 method, and 90% were detected with LJ medium. When contaminated specimens were excluded from the analysis, the LRPs detected 92% (54 of 59) of the cultures. The median time to detection of bacteria was 7 days with both the LRPs and the MGIT 960 method. LRP detection of growth in the presence of p-nitro-α-acetylamino-β-hydroxypropiophenone (NAP) was used for selective identification of M. tuberculosis complex (MTC) and compared to identification with BACTEC 460. Using the LRP NAP test, 47 (94%) out of 50 isolates were correctly identified as tuberculosis complex. The accuracy and speed of LRP antibiotic susceptibility testing with rifampin, streptomycin, isoniazid, and ethambutol were compared to those of the BACTEC 460 method, and discrepant results were checked by the conventional proportion method. In total, 50 MTC isolates were tested. The overall agreement between the LRP and BACTEC 460 results was 98.5%. The median LRP-based susceptibility turnaround time was 2 days (range, 2 to 4 days) compared to 10.5 days (range, 7 to 16 days) by the BACTEC 460 method. Phage resistance was not detected in any of the 243 MTC isolates tested. Mycobacteriophage-based approaches to tuberculosis diagnostics can be implemented in clinical laboratories with sensitivity, specificity, and rapidity that compare favorably with those of the MGIT 960 and BACTEC 460 methods. The phages currently provide the fastest phenotypic assay for susceptibility testing.

AB - The utility of luciferase reporter mycobacteriophages (LRPs) for detection, identification, and antibiotic susceptibility testing of Mycobacterium tuberculosis was prospectively evaluated in a clinical microbiology laboratory in Mexico City, Mexico. Five hundred twenty-three consecutive sputum samples submitted to the laboratory during a 5-month period were included in this study. These specimens were cultivated in Middle-brook 7H9 (MADC), MGIT, and Löwenstein-Jensen (LJ) media. Of the 71 mycobacterial isolates recovered with any of the three media, 76% were detected with the LRPs, 97% were detected with the MGIT 960 method, and 90% were detected with LJ medium. When contaminated specimens were excluded from the analysis, the LRPs detected 92% (54 of 59) of the cultures. The median time to detection of bacteria was 7 days with both the LRPs and the MGIT 960 method. LRP detection of growth in the presence of p-nitro-α-acetylamino-β-hydroxypropiophenone (NAP) was used for selective identification of M. tuberculosis complex (MTC) and compared to identification with BACTEC 460. Using the LRP NAP test, 47 (94%) out of 50 isolates were correctly identified as tuberculosis complex. The accuracy and speed of LRP antibiotic susceptibility testing with rifampin, streptomycin, isoniazid, and ethambutol were compared to those of the BACTEC 460 method, and discrepant results were checked by the conventional proportion method. In total, 50 MTC isolates were tested. The overall agreement between the LRP and BACTEC 460 results was 98.5%. The median LRP-based susceptibility turnaround time was 2 days (range, 2 to 4 days) compared to 10.5 days (range, 7 to 16 days) by the BACTEC 460 method. Phage resistance was not detected in any of the 243 MTC isolates tested. Mycobacteriophage-based approaches to tuberculosis diagnostics can be implemented in clinical laboratories with sensitivity, specificity, and rapidity that compare favorably with those of the MGIT 960 and BACTEC 460 methods. The phages currently provide the fastest phenotypic assay for susceptibility testing.

UR - http://www.scopus.com/inward/record.url?scp=0034753254&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034753254&partnerID=8YFLogxK

U2 - 10.1128/JCM.39.11.3883-3888.2001

DO - 10.1128/JCM.39.11.3883-3888.2001

M3 - Article

C2 - 11682502

AN - SCOPUS:0034753254

VL - 39

SP - 3883

EP - 3888

JO - Journal of Clinical Microbiology

JF - Journal of Clinical Microbiology

SN - 0095-1137

IS - 11

ER -