Comparison of real-time, quantitative PCR with molecular beacons to nested PCR and culture methods for detection of Mycobacterium avium subsp. paratuberculosis in bovine fecal samples.
An automated PCR with fluorescent probes (molecular beacons) detected Mycobacterium avium subsp. paratuberculosis in bovine feces. When the PCR was compared with culture in testing 41 fecal samples, kappa scores of 0.94 to 0.96, a sensitivity of 93 to 96%, and a specificity of 92% were obtained. Results were quantitated by using a standard curve derived from a plasmid containing IS900. A minimum quantity of 1.7 x 10(-4) pg of DNA, correlating to 1 to 8 CFU, was detected.
Fang
Ying
Y
Animal Disease Research and Diagnostic Laboratory, South Dakota State University, Brookings, South Dakota 57007, USA.
Wu
Wai-Hong
WH
Pepper
Jessica L
JL
Larsen
Jill L
JL
Marras
Salvatore A E
SA
Nelson
Eric A
EA
Epperson
William B
WB
Christopher-Hennings
Jane
J
D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000382:microbiology; D003470:Culture Media; D005243:Feces / Q000382:microbiology*; D015335:Molecular Probes; D016927:Mycobacterium avium subsp. paratuberculosis / Q000235:genetics / Q000302:isolation & purification*; D010283:Paratuberculosis / Q000175:diagnosis* / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D003470:Culture Media; D015335:Molecular Probes