phi(2)GFP10, a High-Intensity Fluorophage, Enables Detection and Rapid Drug Susceptibility Testing of Mycobacterium tuberculosis Directly from Sputum Samples

作者：Jain Paras; Hartman Travis E; Eisenberg Nell; O'Donnell Max R; Kriakov Jordan; Govender Karnishree; Makume Mantha; Thaler David S; Hatfull Graham F; Sturm A Willem; Larsen Michelle H; Moodley Preshnie; Jacobs William R Jr^*

来源：Journal of Clinical Microbiology, 2012, 50(4): 1362-1369.

DOI：10.1128/JCM.06192-11

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 (phi(2)GFP10) that for the first time allows direct visualization of Mycobacterium tuberculosis in clinical sputum samples. Engineered features distinguishing phi(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. phi(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 phi(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 phi(2)GFP10. Fluorophage phi(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.

• 出版日期2012-4