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Journal of Clinical Microbiology, January 2002, p. 287-291, Vol. 40, No. 1
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.1.287-291.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

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,1 Wai-Hong Wu,1 Jessica L. Pepper,1 Jill L. Larsen,1 Salvatore A. E. Marras,2 Eric. A. Nelson,1 William B. Epperson,1 and Jane Christopher-Hennings1*

Animal Disease Research and Diagnostic Laboratory, South Dakota State University, Brookings, South Dakota 57007,1 Department of Molecular Genetics, Public Health Research Institute, New York, New York 100162

Received 5 July 2001/ Returned for modification 18 September 2001/ Accepted 14 October 2001

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.

* Corresponding author. Mailing address: Animal Disease Research and Diagnostic Laboratory, South Dakota State University, Brookings, SD 57007. Phone: (605) 688-5171. Fax: (605) 688-6003. E-mail: Jane_Christopher-Hennings{at}sdstate.edu.

Journal of Clinical Microbiology, January 2002, p. 287-291, Vol. 40, No. 1
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.1.287-291.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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