Identification and sub-typing of Mycobacterium avium subsp. paratuberculosis and Mycobacterium avium subsp. avium by randomly amplified polymorphic DNA

A commercially available kit consisting of twenty 10-mer random primers was evaluated to allow selection of a suitable primer that would permit identification and sub-typing of Mycobacterium avium subsp. paratuberculosis and Mycobacterium avium subsp. avium by randomly amplified polymorphic DNA (RAP...

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Bibliographic Details
Published in:Veterinary microbiology 2001-04, Vol.79 (3), p.275-284
Main Authors: Pillai, Shreekumar R, Jayarao, Bhushan M, Gummo, Jennifer D, Hue, Eric C, Tiwari, Deepanker, Stabel, Judith R, Whitlock, Robert H
Format: Article
Language:English
Subjects:
Animals
Bacterial diseases
Bacteriological methods and techniques used in bacteriology
Bacteriology
Biological and medical sciences
Cattle
Cattle Diseases - diagnosis
Cattle Diseases - microbiology
DNA Primers - chemistry
DNA, Bacterial - chemistry
DNA, Bacterial - isolation & purification
Electrophoresis, Agar Gel - veterinary
Fundamental and applied biological sciences. Psychology
Goat Diseases - diagnosis
Goat Diseases - microbiology
Goats
Human bacterial diseases
Humans
Identification
Image Processing, Computer-Assisted
Infectious diseases
Medical sciences
Microbiology
Mycobacterium avium
Mycobacterium avium subsp. paratuberculosis - chemistry
Mycobacterium avium subsp. paratuberculosis - classification
Mycobacterium avium subsp. paratuberculosis - genetics
Mycobacterium paratuberculosis
Paratuberculosis - microbiology
Random Amplified Polymorphic DNA Technique - methods
Random Amplified Polymorphic DNA Technique - veterinary
RAPD
Sheep
Sheep Diseases - diagnosis
Sheep Diseases - microbiology
Sub-typing
Tuberculosis and atypical mycobacterial infections
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Summary:A commercially available kit consisting of twenty 10-mer random primers was evaluated to allow selection of a suitable primer that would permit identification and sub-typing of Mycobacterium avium subsp. paratuberculosis and Mycobacterium avium subsp. avium by randomly amplified polymorphic DNA (RAPD). A primer OPE-20 (5′-AAC-GGT-GAC-C-3′) was identified to be the most suitable primer when tested with four ATCC reference strains of M. paratuberculosis and eight well characterized field strains each of M. paratuberculosis and M. avium. Primer OPE-20 was further tested for its ability to identify and subtype 200 field isolates of M. paratuberculosis. The fingerprint patterns of M. paratuberculosis ( n=212) consisted of five unique common fragments (620, 450, 310, 230, 180 bp) and nine variable fragments resulting in six distinct genotypes. The DNA fingerprints of M. avium ( n=8) consisted of a single common fragment of 620 bp, and 15 variable fragments resulting in six different genotypes. The cattle, human and goat isolates of M. paratuberculosis were genetically similar, but a sheep isolate had a different RAPD profile as compared to RAPD profiles from other species. RAPD was observed to be a rapid, reproducible and reliable technique for identification and sub-typing of M. paratuberculosis.
ISSN:0378-1135
1873-2542
DOI:10.1016/S0378-1135(00)00358-8