Document Detail

Identification of diagnostic proteins in Mycobacterium avium subspecies paratuberculosis by a whole genome analysis approach.
MedLine Citation:
PMID:  16957356     Owner:  NLM     Status:  MEDLINE    
Mycobacterium avium subspecies paratuberculosis (M. paratuberculosis) is an economically significant veterinary pathogen that causes Johne's disease in cattle and sheep. There is a critical need for improved diagnostic tests to detect M. paratuberculosis infection in these animals. As with many other animal diseases, efforts need to be concentrated on the development of simple, rapid, noninvasive tests that can be performed by veterinarians or animal producers without expensive laboratory equipment. With the genome sequence of M. paratuberculosis now complete, we have taken a different strategy to identify novel proteins that are present uniquely in M. paratuberculosis and are antigenic in the context of infected cattle. Through a whole genome comparison of M. paratuberculosis with other sequenced mycobacterial genomes, we identified a collection of more than 90 genes that are present uniquely in M. paratuberculosis. This list has been further trimmed to 39 after amplification using polymerase chain reaction of unique genes using the genomic deoxyribonucleic acid template from several mycobacterial species and isolates. A selection of the remaining genes has been cloned and expressed in Escherichia coli and purified by affinity chromatography. Successfully purified proteins were analyzed using sera from rabbits immunized with M. paratuberculosis. Furthermore, to identify antigens in the context of disease, sera from cattle with Johne's disease as well as healthy control cattle are used in immunoassays. Using this methodology, we identified the first protein antigens specific to M. paratuberculosis.
John P Bannantine; Michael L Paustian
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Publication Detail:
Type:  Evaluation Studies; Journal Article    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  345     ISSN:  1064-3745     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2006  
Date Detail:
Created Date:  2006-09-07     Completed Date:  2006-10-26     Revised Date:  2007-11-15    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  185-96     Citation Subset:  IM    
Bacterial Diseases of Liverstock Research Unit, National Animal Disease Center, USDA-ARS, Ames, IA, USA.
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MeSH Terms
Antigens, Bacterial / diagnostic use*,  genetics,  immunology
Bacterial Proteins / diagnostic use*,  genetics,  immunology
DNA, Bacterial / analysis
Genome, Bacterial*
Mycobacterium avium subsp. paratuberculosis / classification,  genetics,  immunology*,  isolation & purification
Paratuberculosis / diagnosis*,  microbiology
Sequence Analysis, DNA
Reg. No./Substance:
0/Antigens, Bacterial; 0/Bacterial Proteins; 0/DNA, Bacterial

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