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

Ying Fang; Wai-Hong Wu; Jessica L Pepper; Jill L Larsen; Salvatore A E Marras; Eric A Nelson; William B Epperson; Jane Christopher-Hennings

doi:10.1128/JCM.40.1.287-291.2002

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

J Clin Microbiol. 2002 Jan;40(1):287-91. doi: 10.1128/JCM.40.1.287-291.2002.

Authors

Ying Fang¹, Wai-Hong Wu, Jessica L Pepper, Jill L Larsen, Salvatore A E Marras, Eric A Nelson, William B Epperson, Jane Christopher-Hennings

Affiliation

¹ Animal Disease Research and Diagnostic Laboratory, South Dakota State University, Brookings, South Dakota 57007, USA.

Abstract

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.

Publication types

Comparative Study
Evaluation Study
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Animals
Cattle
Cattle Diseases / diagnosis*
Cattle Diseases / microbiology
Culture Media
Feces / microbiology*
Molecular Probes
Mycobacterium avium subsp. paratuberculosis / genetics
Mycobacterium avium subsp. paratuberculosis / isolation & purification*
Paratuberculosis / diagnosis*
Paratuberculosis / microbiology
Polymerase Chain Reaction / methods*
Sensitivity and Specificity

Substances

Culture Media
Molecular Probes