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

Banaiee, N and Bobadilla-del-Valle, M and Bardarov S, J and Riska, PF and Small, PM and Ponce-de-Leon, A and Jacobs, J and Hatfull, GF and Sifuentes-Osornio, J (2001) Luciferase reporter mycobacteriophages for detection, identification, and antibiotic susceptibility testing of mycobacterium tuberculosis in Mexico. Journal of Clinical Microbiology, 39 (11). 3883 - 3888. ISSN 0095-1137

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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.


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Details

Item Type: Article
Status: Published
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Banaiee, N
Bobadilla-del-Valle, M
Bardarov S, J
Riska, PF
Small, PM
Ponce-de-Leon, A
Jacobs, J
Hatfull, GFgfh@pitt.eduGFH
Sifuentes-Osornio, J
Date: 14 November 2001
Date Type: Publication
Journal or Publication Title: Journal of Clinical Microbiology
Volume: 39
Number: 11
Page Range: 3883 - 3888
DOI or Unique Handle: 10.1128/jcm.39.11.3883-3888.2001
Schools and Programs: Dietrich School of Arts and Sciences > Biological Sciences
Refereed: Yes
ISSN: 0095-1137
MeSH Headings: Antitubercular Agents--pharmacology; Culture Media; Genes, Reporter; Humans; Luciferases--genetics; Mexico; Microbial Sensitivity Tests; Mycobacteriophages--genetics; Mycobacteriophages--physiology; Mycobacterium tuberculosis--classification; Mycobacterium tuberculosis--drug effects; Mycobacterium tuberculosis--isolation & purification; Mycobacterium tuberculosis--virology; Tuberculosis--microbiology
Other ID: NLM PMC88459
PubMed Central ID: PMC88459
PubMed ID: 11682502
Date Deposited: 29 Oct 2012 20:29
Last Modified: 02 Feb 2019 15:57
URI: http://d-scholarship.pitt.edu/id/eprint/16119

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