ϕ 2 GFP10, a High-Intensity Fluorophage, Enables Detection and Rapid Drug Susceptibility Testing of Mycobacterium tuberculosis Directly from Sputum Samples
| Autor: | Mantha Makume, Preshnie Moodley, A. Willem Sturm, Max R. O'Donnell, William R. Jacobs, Travis Hartman, Jordan Kriakov, Karnishree Govender, Paras Jain, Michelle H. Larsen, David S. Thaler, Graham F. Hatfull, Nell Eisenberg |
|---|---|
| Rok vydání: | 2012 |
| Předmět: |
Microbiology (medical)
Ofloxacin Tuberculosis medicine.drug_class Genetic Vectors Green Fluorescent Proteins Antibiotics Antitubercular Agents Microbial Sensitivity Tests Drug resistance Signal-To-Noise Ratio Biology Microbiology Mycobacterium tuberculosis Genes Reporter Kanamycin Isoniazid medicine Humans Bacteriophages Promoter Regions Genetic Tuberculosis Pulmonary Sputum Mycobacteriology and Aerobic Actinomycetes biology.organism_classification medicine.disease Virology Recombinant Proteins Rifampin medicine.symptom Rifampicin medicine.drug |
| Zdroj: | Journal of Clinical Microbiology. 50:1362-1369 |
| ISSN: | 1098-660X 0095-1137 |
| DOI: | 10.1128/jcm.06192-11 |
| Popis: | 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 2 GFP10) that for the first time allows direct visualization of Mycobacterium tuberculosis in clinical sputum samples. Engineered features distinguishing ϕ 2 GFP10 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 2 GFP10 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 2 GFP10 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 2 GFP10. Fluorophage ϕ 2 GFP10 has potential for clinical development as a rapid, sensitive, and inexpensive point-of-care diagnostic tool for M. tuberculosis infection and for rapid DST. |
| Databáze: | OpenAIRE |
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