Detection of viable Mycobacterium avium subsp. paratuberculosis using luciferase reporter systems.
Plasmid- and phage-based firefly luciferase reporter constructs were evaluated as rapid detection systems for viable Mycobacterium avium subsp. paratuberculosis (MAP). A MAP strain bearing a luciferase-encoding plasmid was detectable at 100 cells/mL in skim milk and 1000 cells/mL in whole milk. Three luciferase-encoding mycobacteriophage were evaluated for detection of wild-type MAP. The best of these, phAE85, allowed detection of >1000 cells/mL within 24-48 h. Membrane filtration did not improve the sensitivity of detection for either plasmid or phage reporters. Luciferase reporters show promise for rapid detection of viable MAP.
Sasahara
Kyle C
KC
Food Safety Laboratory, Department of Food Science, Cornell University, Ithaca, New York 14853, USA.
Gray
Michael J
MJ
Shin
Sang J
SJ
Boor
Kathryn J
KJ
D000818:Animals; D002417:Cattle; D015169:Colony Count, Microbial; D005506:Food Contamination / Q000032:analysis*; D005516:Food Microbiology; D049453:Luciferases, Bacterial / Q000235:genetics* / Q000378:metabolism; D008892:Milk / Q000382:microbiology*; D016927:Mycobacterium avium subsp. paratuberculosis / Q000302:isolation & purification*; D010957:Plasmids; D013997:Time Factors
D049453:Luciferases, Bacterial