Comparison of two DNA extractions and nested PCR, real-time PCR, a new commercial PCR assay, and bacterial culture for detection of Mycobacterium avium subsp. paratuberculosis in bovine feces.
In this study, 5 combinations of 2 DNA extractions and 3 polymerase chain reaction (PCR) techniques were compared with culture for the detection of Mycobacterium paratuberculosis directly from bovine feces. These combinations included a new commercial extraction technique combined with a commercial PCR/Southern blot technique, nested PCR (nPCR), or real-time PCR, and a university-developed extraction combined with nPCR or real-time PCR. Four of the 5 combinations had statistically similar sensitivities between 93% and 100% and specificity between 95% and 100%, when compared with culture results from 63 bovine fecal samples. These results indicated that using a commercial extraction with a commercial PCR/Southern blot, nPCR, or real-time PCR, or a university-developed extraction with real-time PCR would result in similar sensitivities to culture for the identification of M. paratuberculosis from bovine feces and are valid alternatives to culture.
Christopher-Hennings
Jane
J
Animal Disease Research and Diagnostic Laboratory, South Dakota State University, Brookings, SD 57007, USA.
Dammen
Matthew A
MA
Weeks
Shelleen R
SR
Epperson
William B
WB
Singh
Shri N
SN
Steinlicht
Gina L
GL
Fang
Ying
Y
Skaare
Jessica L
JL
Larsen
Jill L
JL
Payeur
Janet B
JB
Nelson
Eric A
EA
D000818:Animals; D001431:Bacteriological Techniques / Q000379:methods; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000382:microbiology*; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification*; D005243:Feces / Q000382:microbiology*; D016927:Mycobacterium avium subsp. paratuberculosis / Q000302:isolation & purification*; D010283:Paratuberculosis / Q000175:diagnosis* / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D004269:DNA, Bacterial