Detection of Mycobacterium paratubeculosis in feces is complicated by the slowdivision and the lipid composition of the cell wall of the bacterium. In this study, we compared a rapid and sensitive method for direct detection of M. paratuberculosis with anti-paratuberculosis chicken IgY-conjugated magnetic nanoparticles versus commercially available micro-sized paramagnetic beads. Tenfold serial dilutions of a standard M. paratuberculosis strain and clinical isolation strains were spiked in pooled stool and detected by using antibody-conjugated magnetic nanoparticles or paramagnetic beads, and confirmed by polymerase chain reaction. The binding ratio of antibody-nanoparticles was 96.44 ± 0.77%, and the detection limit was 20 M. paratuberculosis cells per gram of feces. Also, magnetic nanoparticles showed significantly intensive results than paramagnetic beads in dose-effects evaluation. The magnetic nanoparticles were also cost-effective. Antibody-conjugated magnetic nanoparticles did not react with other bacteria, thus providing specific immunomagnetic separation. We conclude that the antibody-conjugated magnetic nanoparticles are more effective than paramagnetic beads for low-cost, direct detection of M. paratuberculosis in cattle feces; in addition, confirmation time was reduced from 12−16 weeks to 2−3 days. This antibody-based immunocapture polymerase chain reaction method is expected to provide a platform of rapid screening of M. paratuberculosis in fecal samples in one step.
|Number of pages||9|
|Journal||International Journal of Applied Research in Veterinary Medicine|
|Publication status||Published - 2016 Jan 1|
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