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.
References
1.
BannantineJPZhangQLiLL: 2003, Genomic homogeneity between Mycobacterium avium subsp. avium and Mycobacterium avium subsp. paratuberculosis belies their divergent growth rates. BMC Microbiol3:10–19.
2.
BullTJHermon-TaylorJPavlikII: 2000, Characterization of IS900 loci in Mycobacterium avium subsp. paratuberculosis and development of multiplex PCR typing. Microbiology146:3285.
3.
ClarkstonWKPrestiMEPetersenPF: 1998, Role of Mycobacterium paratuberculosis in Crohn's disease: a prospective, controlled study using polymerase chain reaction. Dis Colon Rectum41:195–199.
4.
CollinsDMGabricDMDe LisleGW: 1989, Identification of a repetitive DNA sequence specific to Mycobacterium paratuberculosis. FEMS Microbiol Lett51:175–178.
5.
CollinsMT: 1996, Diagnosis of paratuberculosis. Vet Clin North Am Food Anim Pract12:357–371.
6.
CousinsDVWhittingtonRMarshI: 1999, Mycobacteria distinct from Mycobacterium avium subsp. paratuberculosis isolated from the faeces of ruminants possess IS900-like sequences detectable IS900 polymerase chain reaction: implications for diagnosis. Mol Cell Probes13:431–442.
7.
DamatoJJKnisleyCCollinsMT: 1987, Characterization of Mycobacterium paratuberculosis by gas-liquid and thin-layer chromatography and rapid demonstration of mycobactin dependence using radiometric methods. J Clin Microbiol25:2380–2383.
8.
Dell'IsolaBPoyartCGouletO: 1994, Detection of Mycobacterium paratuberculosis by polymerase chain reaction in children with Crohn's disease. J Infect Dis169:449–451.
9.
DumonceauJMVan GossumAAdlerM: 1996, No Mycobacterium paratuberculosis found in Crohn's disease using polymerase chain reaction. Dig Dis Sci41:421–426.
10.
DundeeLGrantIRBallHJ: 2001, Comparative evaluation of four decontamination protocols for the isolation of Mycobacterium avium subsp. paratuberculosis from milk. Lett Appl Microbiol33:173–177.
11.
ElsaghierAPranteraCMorenoC: 1992, Antibodies to Mycobacterium paratuberculosis-specific protein antigens in Crohn's disease. Clin Exp Immunol90:503–508.
EriksISMunckKTBesserTE: 1996, Rapid differentiation of Mycobacterium avium and M. paratuberculosis by PCR and restriction enzyme analysis. J Clin Microbiol34:734–737.
14.
GreenEPTizardMLMossMT: 1989, Sequence and characteristics of IS900, an insertion element identified in a human Crohn's disease isolate of Mycobacterium paratuberculosis. Nucleic Acids Res17:9063–9073.
15.
HarrisJELammerdingAM: 2001, Crohn's disease and Mycobacterium avium subsp. paratuberculosis: current issues. J Food Prot64:2103–2110.
16.
Hermon-TaylorJ: 1993, Causation of Crohn's disease: the impact of clusters. Gastroenterology104:643–646.
17.
Hermon-TaylorJBullT: 2002, Crohn's disease caused by Mycobacterium avium subspecies paratuberculosis: a public health tragedy whose resolution is long overdue. J Med Microbiol51:3–6.
HurleySSSplitterGAWelchRA: 1989, Development of a diagnostic test for Johne's disease using a DNA hybridization probe. J Clin Microbiol27:1582–1587.
20.
JorgensenJB: 1982, An improved medium for culture of Mycobacterium paratuberculosis from bovine faeces. Acta Vet Scand23:325–335.
21.
JusteRAMarcoJCSaez de OcarizC: 1991, Comparison of different media for the isolation of small ruminant strains of Mycobacterium paratuberculosis. Vet Microbiol28:385–390.
22.
KalisCHHesselinkJWRusschenEW: 1999, Factors influencing the isolation of Mycobacterium avium subsp. paratuberculosis from bovine fecal samples. J Vet Diagn Invest11: 345–351.
23.
KimSGShinSJJacobsonRH: 2002, Development and application of quantitative polymerase chain reaction assay based on the ABI 7700 system (TaqMan) for detection and quantification of Mycobacterium avium subsp. paratuberculosis. J Vet Diagn Invest14:126–131.
24.
KunzeZMWallSAppelbergR: 1991, IS901, a new member of a widespread class of atypical insertion sequences, is associated with pathogenicity in Mycobacterium avium. Mol Microbiol5:2265–2272.
25.
OttSLWellsSJWagnerBA: 1999, Herd-level economic losses associated with Johne's disease on US dairy operations. Prev Vet Med40:179–192.
26.
RowbothamDSMapstoneNPTrejdosiewiczLK: 1995, Mycobacterium paratuberculosis DNA not detected in Crohn's disease tissue by fluorescent polymerase chain reaction. Gut37:660–667.
27.
RyanPBennettMWAaronsS: 2002, PCR detection of Mycobacterium paratuberculosis in Crohn's disease granulomas isolated by laser capture microdissection. Gut51:665–670.
28.
ShafranIKuglerLEl-ZaatariFA: 2002, Open clinical trial of rifabutin and clarithromycin therapy in Crohn's disease. Dig Liver Dis34:22–28.
29.
ShinSJ: 1989, New methods for reduction in bacterial and fungal contamination from fecal-culture for Mycobacterium paratuberculosis. In: Report of the committee on Johne's disease. Proceedings of the 93rd annual meeting of the US Animal Health Association, Las Vegas, p. 381.
30.
ShinSJKimSGMillerLJ: 2001, Further evaluation of ESP culture system II for detection of Mycobacterium avium subsp. paratuberculosis in bovine fecal samples. American Association of Veterinary Laboratory Diagnosticians, 44th Annual Meeting, p. 39. [Abstr.].
31.
StabelJR: 1997, An improved method for cultivation of Mycobacterium paratuberculosis from bovine fecal samples and comparison to three other methods. J Vet Diagn Invest9:375–380.
32.
StabelJR: 1998, Johne's disease: a hidden threat. J Dairy Sci81:283–288.
33.
StrommengerBStevensonKGerlachGF: 2001, Isolation and diagnostic potential of ISMav2, a novel insertion sequence-like element from Mycobacterium avium subspecies paratuberculosis. FEMS Microbiol Lett196:31–37.
34.
ThayerWR: 1992, The use of antimycobacterial agents in Crohn's disease. J Clin Gastroenterol15:5–7.
35.
Van KruiningenHJ: 1999, Lack of support for a common etiology in Johne's disease of animals and Crohn's disease in humans. Inflamm Bowel Dis5:183–191.
36.
VaryPHAndersenPRGreenE: 1990, Use of highly specific DNA probes and the polymerase chain reaction to detect Mycobacterium paratuberculosis in Johne's disease. J Clin Microbiol28:933–937.
37.
van SoolingenDHermansPW: 1995, Epidemiology of tuberculosis by DNA fingerprinting. Eur Respir J Suppl20:649s–656s.
38.
WellsSJWagnerBA: 2000, Herd-level risk factors for infection with Mycobacterium paratuberculosis in US dairies and association between familiarity of the herd manager with the disease or prior diagnosis of the disease in that herd and use of preventive measures. J Am Vet Med Assoc216:1450–1457.
39.
WhippleDLCallihanDRJarnaginJL: 1991, Cultivation of Mycobacterium paratuberculosis from bovine fecal specimens and a suggested standardized procedure. J Vet Diagn Invest3:368–373.
40.
WhittingtonRJFellSWalkerD: 2000, Use of pooled fecal culture for sensitive and economic detection of Mycobacterium avium subsp. paratuberculosis infection in flocks of sheep. J Clin Microbiol38:2550–2556.
