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Mycobacterium paratuberculosis cultu...

Cho, Donghee.

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Mycobacterium paratuberculosis culture filtrate antigens.

Record Type:	Language materials, printed : Monograph/item
Title/Author:	Mycobacterium paratuberculosis culture filtrate antigens./
Author:	Cho, Donghee.
Description:	193 p.
Notes:	Adviser: Michael T. Collins.
Contained By:	Dissertation Abstracts International67-09B.
Subject:	Biology, Veterinary Science. -
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3234572
ISBN:	9780542885198

Mycobacterium paratuberculosis culture filtrate antigens.
Cho, Donghee.

Mycobacterium paratuberculosis culture filtrate antigens. - 193 p.

Adviser: Michael T. Collins.

Thesis (Ph.D.)--The University of Wisconsin - Madison, 2006.

Control of bovine paratuberculosis, caused by Mycobacterium paratuberculosis, requires a combination of herd management changes to limit infection transmission and identification of the infection source, i.e., adult cattle shedding the pathogen. Diagnostic tests based on detection of antibodies to M. paratuberculosis in serum or milk by ELISA are favored for their low cost and high assay throughput. Existing paratuberculosis ELISAs are based on antigens from M. paratuberculosis cellular extracts (CE) and suffer from low diagnostic sensitivity, e.g., &sim;30%. Recently, the field of mycobacteria diagnostics has shifted its attention to secreted proteins, recognizing that culture filtrates (CF) contain important immunodominant antigens. The CF of M. paratuberculosis was the focus of this thesis. From among roughly 200 CF proteins, specific proteins and peptides potentially useful for serodiagnosis were identified by two-dimensional electrophoresis (2-DE) immunoblots, and MALDI-MS. A total of 14 proteins were identified by mass spectrometry and BLAST search as ModD, PepA, ArgJ, CobT, Antigen 85C and 9 hypothetical proteins. All 14 ORFs were expressed in E.coli BL21(DE3). Sera from cattle with subclinical infections reacted to five of these cloned proteins including ModD, Antigen 85C, PepA, MAP1693c, and MAP2168c, with reactions to ModD being the strongest. A ModD ELISA, however, showed low specificity (high background with sera from noninfected cattle). Mapping of ModD was done by creating 80 overlapping peptides (14 amino acids) and testing them with antisera from immunized rabbits and sera from subclinical cases of bovine paratuberculosis. Four peptides, located on the C-terminal ends of ModD appear to be M. paratuberculosis-specific epitopes, reacting strongly with rabbit anti-M. paratuberculosis CF and weakly with anti-M. avium CF. Further studies using this immunological approach to antigen and epitope identification and characterization may lead to improved paratuberculosis diagnostics.

ISBN: 9780542885198Subjects--Topical Terms:

1021733
Biology, Veterinary Science.

Mycobacterium paratuberculosis culture filtrate antigens.
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100 1 $a Cho, Donghee. $3 1269253
245 1 0 $a Mycobacterium paratuberculosis culture filtrate antigens.
300 $a 193 p.
500 $a Adviser: Michael T. Collins.
500 $a Source: Dissertation Abstracts International, Volume: 67-09, Section: B, page: 4909.
502 $a Thesis (Ph.D.)--The University of Wisconsin - Madison, 2006.
520 $a Control of bovine paratuberculosis, caused by Mycobacterium paratuberculosis, requires a combination of herd management changes to limit infection transmission and identification of the infection source, i.e., adult cattle shedding the pathogen. Diagnostic tests based on detection of antibodies to M. paratuberculosis in serum or milk by ELISA are favored for their low cost and high assay throughput. Existing paratuberculosis ELISAs are based on antigens from M. paratuberculosis cellular extracts (CE) and suffer from low diagnostic sensitivity, e.g., &sim;30%. Recently, the field of mycobacteria diagnostics has shifted its attention to secreted proteins, recognizing that culture filtrates (CF) contain important immunodominant antigens. The CF of M. paratuberculosis was the focus of this thesis. From among roughly 200 CF proteins, specific proteins and peptides potentially useful for serodiagnosis were identified by two-dimensional electrophoresis (2-DE) immunoblots, and MALDI-MS. A total of 14 proteins were identified by mass spectrometry and BLAST search as ModD, PepA, ArgJ, CobT, Antigen 85C and 9 hypothetical proteins. All 14 ORFs were expressed in E.coli BL21(DE3). Sera from cattle with subclinical infections reacted to five of these cloned proteins including ModD, Antigen 85C, PepA, MAP1693c, and MAP2168c, with reactions to ModD being the strongest. A ModD ELISA, however, showed low specificity (high background with sera from noninfected cattle). Mapping of ModD was done by creating 80 overlapping peptides (14 amino acids) and testing them with antisera from immunized rabbits and sera from subclinical cases of bovine paratuberculosis. Four peptides, located on the C-terminal ends of ModD appear to be M. paratuberculosis-specific epitopes, reacting strongly with rabbit anti-M. paratuberculosis CF and weakly with anti-M. avium CF. Further studies using this immunological approach to antigen and epitope identification and characterization may lead to improved paratuberculosis diagnostics.
590 $a School code: 0262.
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710 2 $a The University of Wisconsin - Madison. $3 626640
773 0 $t Dissertation Abstracts International $g 67-09B.
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791 $a Ph.D.
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