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.
Food Safety Laboratory, Department of Food Science, Cornell University, Ithaca, New York 14853, USA.
Kyle C
KC
Sasahara
Michael J
MJ
Gray
Sang J
SJ
Shin
Kathryn J
KJ
Boor
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