Production of and Applications for a Polyclonal IgY Diagnostic Reagent Specific for Mycobacterium avium subsp. paratuberculosis

Sung Jae Shin; Seung-Sub Lee; Elizabeth J. B. Manning; Michael T. Collins

doi:10.1007/s12275-009-0052-7

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Research Support, Non-U.S. Gov't
Production of and Applications for a Polyclonal IgY Diagnostic Reagent Specific for Mycobacterium avium subsp. paratuberculosis: Sung Jae Shin ¹, Seung-Sub Lee ¹, Elizabeth J. B. Manning ², Michael T. Collins ²; Journal of Microbiology 2009;47(5):600-609.
DOI: https://doi.org/10.1007/s12275-009-0052-7
Published online: October 24, 2009

¹Department of Microbiology and Infection Signaling Network Research Center, Research Institute for Medical Sciences, College of Medicine, Chungnam National University, Daejeon 301-747, Republic of Korea, ²Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, 2015 Linden Drive, Madison, WI 53706-1102, USA

Corresponding author: Elizabeth J. B. Manning , Tel: 1-608-265-4958,

Received: 23 February 2009 • Accepted: 12 May 2009

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Abstract

Antibodies specific to the cell surface antigens of Mycobacterium avium subsp. paratuberculosis (MAP) have multiple useful applications, e.g. organism detection, immunoconcentration, and cell visualization. The aim of this study was to produce and compare polyclonal antibodies for such research and diagnostic purposes. Three polyclonal antibodies to MAP were produced using sera from immunized rabbits and chickens plus naturally infected cows. Cross-reactive antibodies in each MAP antibody preparation were removed by absorption with heterologous mycobacterial and non-mycobacterial cells. The specificity of each resulting polyclonal antibody preparation was evaluated by ELISA to multiple bacterial cell wall extract antigens. After absorption, chicken anti-MAP IgY had the highest specificity of the three antibody preparations. FITC-labeled anti-MAP IgY was used to effectively locate MAP in macrophages 12 h post-infection. Also, immuno- magnetic beads coated with anti-MAP IgY enhanced recovery of MAP from bacterial suspensions in comparison with non-antibody coated beads. Anti-MAP IgY provides a novel new reagent with broad diagnostic and research applications requiring specific concentration, detection, and quantification of MAP.

Keywords: M. paratuberculosis;polyclonal antibodies for diagnostic purposes;absorption;anti-MAP IgY

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Production of and Applications for a Polyclonal IgY Diagnostic Reagent Specific for Mycobacterium avium subsp. paratuberculosis

J. Microbiol. 2009;47(5):600-609. Published online October 24, 2009

DOI: https://doi.org/10.1007/s12275-009-0052-7

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