TY - JOUR
T1 - Conditionally replicating luciferase reporter phages
T2 - Improved sensitivity for rapid detection and assessment of drug susceptibility of Mycobacterium tuberculosis
AU - Carrière, Christian
AU - Riska, Paul F.
AU - Zimhony, Oren
AU - Kriakov, Jordan
AU - Bardarov, Stoyan
AU - Burns, Judah
AU - Chan, John
AU - Jacobs, William R.
PY - 1997/12
Y1 - 1997/12
N2 - TM4 is a lytic mycobacteriophage which infects mycobacteria of clinical importance. A luciferase reporter phage, phAE40, has been constructed from TM4 and was previously shown to be useful for the rapid detection and drug susceptibility testing of Mycobacterium tuberculosis. However, the lyric nature of the phage results in a loss of detectable light output and limits the sensitivity of detection. We describe several strategies aimed at improving the luciferase activity generated by TM4 luciferase phages, including (i) varying the position of the luciferase gene in the phage genome, (ii) isolating host-range mutants of the phage, and (iii) introducing temperature-sensitive mutations in the phage such that it will not replicate at the infecting temperature. Several new phages generated by these methods show increased intensity of luciferase production compared to the first- generation reporter phage phAE40, and one phage, phAE88, also demonstrates an enhanced duration of luciferase activity. This has allowed the detection of as few as 120 BCG cells and the determination of drug susceptibilities of M. tuberculosis in as little as 1 day.
AB - TM4 is a lytic mycobacteriophage which infects mycobacteria of clinical importance. A luciferase reporter phage, phAE40, has been constructed from TM4 and was previously shown to be useful for the rapid detection and drug susceptibility testing of Mycobacterium tuberculosis. However, the lyric nature of the phage results in a loss of detectable light output and limits the sensitivity of detection. We describe several strategies aimed at improving the luciferase activity generated by TM4 luciferase phages, including (i) varying the position of the luciferase gene in the phage genome, (ii) isolating host-range mutants of the phage, and (iii) introducing temperature-sensitive mutations in the phage such that it will not replicate at the infecting temperature. Several new phages generated by these methods show increased intensity of luciferase production compared to the first- generation reporter phage phAE40, and one phage, phAE88, also demonstrates an enhanced duration of luciferase activity. This has allowed the detection of as few as 120 BCG cells and the determination of drug susceptibilities of M. tuberculosis in as little as 1 day.
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U2 - 10.1128/jcm.35.12.3232-3239.1997
DO - 10.1128/jcm.35.12.3232-3239.1997
M3 - Article
C2 - 9399525
AN - SCOPUS:1842417174
SN - 0095-1137
VL - 35
SP - 3232
EP - 3239
JO - Journal of Clinical Microbiology
JF - Journal of Clinical Microbiology
IS - 12
ER -