Development of a polymerase chain reaction test to confirm Mycobacterium avium subsp. paratuberculosis in culture.
A polymerase chain reaction (PCR) assay for confirmation of Mycobacterium avium subsp. paratuberculosis was developed using the primer set derived from ISMav2. The PCR product was 494 base pairs (bp) and could be digested with ClaI, which produced 311- and 183-bp fragments. No amplification of 494-bp DNA fragment was detected from DNA of other Mycobacterium spp., including Mycobacterium avium complex, other bacteria, including Escherichia coli, Pseudomonas aeruginosa, Actinobacillus pleuropneumoniae, Leptospira interrogans serovar pomona, Corynebacterium pseudotuberculosis, Salmonella typhimurium, Borrelia burgdorferi, and Staphylococcus aureus, and the Scedosporium sp. This PCR assay could detect 5-8 genome equivalents.
Shin
Sung Jae
SJ
Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA.
Chang
Yung-Fu
YF
Huang
Cathy
C
Zhu
Jiaqian
J
Huang
Lester
L
Yoo
Han Sang
HS
Shin
Kwang-Soon
KS
Stehman
Susan
S
Shin
Sang J
SJ
Torres
Alfonso
A
D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000382:microbiology*; D004251:DNA Transposable Elements / Q000235:genetics; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D005243:Feces / Q000382:microbiology; D005260:Female; D009164:Mycobacterium Infections / Q000382:microbiology / Q000662:veterinary*; D016927:Mycobacterium avium subsp. paratuberculosis / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*
D004251:DNA Transposable Elements; D004269:DNA, Bacterial; C032824:ATCGAT-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific