bacterial thymidine kinase as a non-invasive imaging reporter for mycobacterium tuberculosis in live animals细菌胸苷激酶作为非侵入性成像记者结核分枝杆菌的活的动物.pdf

Bacterial Thymidine Kinase as a Non-Invasive Imaging Reporter for Mycobacterium tuberculosis in Live Animals 1,2 1 1,3 4 4 Stephanie L. Davis , Nicholas A. Be , Gyanu Lamichhane , Sridhar Nimmagadda , Martin G. Pomper , William R. Bishai1,3, Sanjay K. Jain1,2* 1 Center for Tuberculosis Research, Johns Hopkins University School of Medicine, Baltimore, Maryland, United States of America, 2 Department of Pediatrics, Johns Hopkins University School of Medicine, Baltimore, Maryland, United States of America, 3 Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland, United States of America, 4 Department of Radiology, Johns Hopkins University School of Medicine, Baltimore, Maryland, United States of America Abstract Background: Bacteria can be selectively imaged in experimentally-infected animals using exogenously administered 1- (29deoxy-29-fluoro-b-D-arabinofuranosyl)-5-[125I]-iodouracil ([125I]-FIAU), a nucleoside analog substrate for bacterial thymidine kinase (TK). Our goal was to use this reporter and develop non-invasive methods to detect and localize Mycobacterium tuberculosis. Methodology/Principal Findings: We engineered a M. tuberculosis strain with chromosomally integrated bacterial TK under the control of hsp60 - a strong constitutive mycobacterial promoter. [125I]FIAU uptake, antimicrobial susceptibilities and in vivo growth characteristics were evaluated for this strain. Using single photon emission computed tomography (SPECT), M. tuberculosis Phsp60 TK strain was evaluated in experimentally-infected BALB/c and C3HeB/