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
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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Item Type: |
Article
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Status: |
Published |
Creators/Authors: |
Creators | Email | Pitt Username | ORCID ![](/images/orcid_id_24x24.png) |
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Jain, P | | | | Hartman, TE | | | | Eisenberg, N | | | | O'Donnell, MR | | | | Kriakov, J | | | | Govender, K | | | | Makume, M | | | | Thaler, DS | | | | Hatfull, GF | gfh@pitt.edu | GFH | | Sturm, AW | | | | Larsen, MH | | | | Moodley, P | | | | Jacobs, WR | | | |
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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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