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φ <sup>2</sup>GFP10, a high-intensity fluorophage, enables detection and rapid drug susceptibility testing of Mycobacterium tuberculosis directly from sputum samples

Jain, P and Hartman, TE and Eisenberg, N and O'Donnell, MR and Kriakov, J and Govender, K and Makume, M and Thaler, DS and Hatfull, GF and Sturm, AW and Larsen, MH and Moodley, P and Jacobs, WR (2012) φ <sup>2</sup>GFP10, a high-intensity fluorophage, enables detection and rapid drug susceptibility testing of Mycobacterium tuberculosis directly from sputum samples. Journal of Clinical Microbiology, 50 (4). 1362 - 1369. ISSN 0095-1137

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Abstract

The difficulty of diagnosing active tuberculosis (TB) and lack of rapid drug susceptibility testing (DST) at the point of care remain critical obstacles to TB control. This report describes a high-intensity mycobacterium-specific- fluorophage (φ 2GFP10) that for the first time allows direct visualization of Mycobacterium tuberculosis in clinical sputum samples. Engineered features distinguishing φ 2GFP10 from previous reporter phages include an improved vector backbone with increased cloning capacity and superior expression of fluorescent reporter genes through use of an efficient phage promoter. φ 2GFP10 produces a 100-fold increase in fluorescence per cell compared to existing reporter phages. DST for isoniazid and oxofloxacin, carried out in cultured samples, was complete within 36 h. Use of φ 2GFP10 detected M. tuberculosis in clinical sputum samples collected from TB patients. DST for rifampin and kanamycin from sputum samples yielded results after 12 h of incubation with φ 2GFP10. Fluorophage φ 2GFP10 has potential for clinical development as a rapid, sensitive, and inexpensive point-of-care diagnostic tool for M. tuberculosis infection and for rapid DST. Copyright © 2012, American Society for Microbiology. All Rights Reserved.

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Details

Item Type: Article
Status: Published
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Jain, P
Hartman, TE
Eisenberg, N
O'Donnell, MR
Kriakov, J
Govender, K
Makume, M
Thaler, DS
Hatfull, GFgfh@pitt.eduGFH
Sturm, AW
Larsen, MH
Moodley, P
Jacobs, WR
Date: 1 April 2012
Date Type: Publication
Journal or Publication Title: Journal of Clinical Microbiology
Volume: 50
Number: 4
Page Range: 1362 - 1369
DOI or Unique Handle: 10.1128/jcm.06192-11
Schools and Programs: Dietrich School of Arts and Sciences > Biological Sciences
Refereed: Yes
ISSN: 0095-1137
MeSH Headings: Antitubercular Agents--pharmacology; Bacteriophages--genetics; Genes, Reporter; Genetic Vectors; Green Fluorescent Proteins--biosynthesis; Green Fluorescent Proteins--genetics; Humans; Isoniazid--pharmacology; Kanamycin--pharmacology; Microbial Sensitivity Tests--methods; Mycobacterium tuberculosis--drug effects; Mycobacterium tuberculosis--metabolism; Mycobacterium tuberculosis--virology; Ofloxacin--pharmacology; Promoter Regions, Genetic; Recombinant Proteins--biosynthesis; Recombinant Proteins--genetics; Rifampin--pharmacology; Signal-To-Noise Ratio; Sputum--microbiology; Tuberculosis, Pulmonary--diagnosis; Tuberculosis, Pulmonary--microbiology
Other ID: NLM PMC3318544
PubMed Central ID: PMC3318544
PubMed ID: 22278833
Date Deposited: 29 Nov 2012 21:04
Last Modified: 29 Sep 2022 04:55
URI: http://d-scholarship.pitt.edu/id/eprint/16303

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