Production of and applications for a polyclonal IgY diagnostic reagent specific for mycobacterium avium subsp. paratuberculosis

SungJae Shin, Seung Sub Lee, Elizabeth J.B. Manning, Michael T. Collins

Research output: Contribution to journalArticle

10 Citations (Scopus)

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-la-beled anti-MAP IgY was used to effectively locate MAP in macrophages 12 h post-infection. Also, immunomagnetic 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.

Original languageEnglish
Pages (from-to)600-609
Number of pages10
JournalJournal of Microbiology
Volume47
Issue number5
DOIs
Publication statusPublished - 2009 Oct 1

Fingerprint

Paratuberculosis
Mycobacterium avium
Antibodies
Chickens
Antibody Specificity
Fluorescein-5-isothiocyanate
Surface Antigens
Cell Extracts
Research
Cell Wall
IgY
Suspensions
Enzyme-Linked Immunosorbent Assay
Macrophages
Rabbits
Antigens
Infection
Serum

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Applied Microbiology and Biotechnology

Cite this

Shin, SungJae ; Lee, Seung Sub ; Manning, Elizabeth J.B. ; Collins, Michael T. / Production of and applications for a polyclonal IgY diagnostic reagent specific for mycobacterium avium subsp. paratuberculosis. In: Journal of Microbiology. 2009 ; Vol. 47, No. 5. pp. 600-609.
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Production of and applications for a polyclonal IgY diagnostic reagent specific for mycobacterium avium subsp. paratuberculosis. / Shin, SungJae; Lee, Seung Sub; Manning, Elizabeth J.B.; Collins, Michael T.

In: Journal of Microbiology, Vol. 47, No. 5, 01.10.2009, p. 600-609.

Research output: Contribution to journalArticle

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