Comparison of Two Immunomagnetic Separation Methods for Detection of Mycobacterium avium subsp Paratuberculosis in Bovine Feces
- Authors
- Kim, Narae; Kang, Hwan-Goo; Jang, Yun-Ho; Ryoo, Soyoon; Lee, Hyuk-Mi; Shin, Sung Jae; Jang, Boseung; Cho, Hyeri; Jung, Suk Chan; Park, Yong Ho; Lim, Young-Hee; Kim, Jae-Myung
- Issue Date
- 2016
- Publisher
- VETERINARY SOLUTIONS LLC
- Keywords
- Immunomagnetic separation; IS900; Johne' s disease; Magnetic nanoparticle; Mycobacterium avium subsp paratuberculosis
- Citation
- INTERNATIONAL JOURNAL OF APPLIED RESEARCH IN VETERINARY MEDICINE, v.14, no.1, pp.96 - 104
- Indexed
- SCIE
SCOPUS
- Journal Title
- INTERNATIONAL JOURNAL OF APPLIED RESEARCH IN VETERINARY MEDICINE
- Volume
- 14
- Number
- 1
- Start Page
- 96
- End Page
- 104
- URI
- https://scholar.korea.ac.kr/handle/2021.sw.korea/90257
- ISSN
- 1542-2666
- Abstract
- 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.
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Collections - College of Health Sciences > School of Biosystems and Biomedical Sciences > 1. Journal Articles
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