DNA fingerprinting of Australian isolates of <i>Mycobacterium avium subsp paratuberculosis</i> using IS<i>900</i> RFLP

Cousins, DV; Williams, S.N.; Hope, A. F.; Eamens, G. J.

doi:10.1111/j.1751-0813.2000.tb10590.x

Cited by 54 publications

(49 citation statements)

References 22 publications

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“…4). This profile appears to be similar to the IS900-RFLP BstEII S profiles previously described by Collins et al (5), Whittington et al (27), and Cousins et al (7). The pigmented isolates had identical IS900-RFLP PvuII profiles (Fig.…”

Section: Resultssupporting

confidence: 63%

“…The pigmented isolates had identical IS900-RFLP PvuII profiles (Fig. 5), which appeared to correspond to PvuII type 6 (described by Cousins et al [7]). In contrast, nonpigmented isolates representative of the different multiplex PFGE profiles were found to have BstEII profiles corresponding to C1, C5, and C17 (defined by Pavlik et al [15]) and two new profiles not previously published (Table 1) and PvuII profiles corresponding to types 1, 2, and 3 (defined by Whipple et al [25]) and one new profile not previously described.…”

Section: Resultsmentioning

confidence: 86%

“…paratuberculosis. In Australia and New Zealand, S strains are the predominant strains isolated from sheep (7,27). However, in Europe, where paratuberculosis is endemic, C strains are more commonly isolated from sheep as well as from cattle and nonruminant hosts (13,16).…”

Section: Discussionmentioning

confidence: 99%

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Molecular Characterization of Pigmented and Nonpigmented Isolates of Mycobacterium avium subsp. paratuberculosis

et al. 2002

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Five pigmented isolates of Mycobacterium avium subsp. paratuberculosis were examined by pulsed-field gel electrophoresis (PFGE), IS900 restriction fragment length polymorphism (IS900-RFLP), and IS1311 polymorphism analysis using PCR. All of the pigmented isolates exhibited one of three distinct PFGE profiles with SnaBI, designated 9, 10, and 11, and with SpeI, designated 7, 8, and 9, which generated three multiplex profiles designated . IS1311-PCR analysis typed all of the pigmented isolates as sheep (S) strains. The genetic relationship between pigmented and nonpigmented isolates was investigated by using multiplex PFGE data from the analysis of both the 5 pigmented isolates and 88 nonpigmented isolates of M. avium subsp. paratuberculosis from a variety of host species and geographic locations. It was possible to classify the isolates into two distinct types designated type I, comprising the pigmented isolates, and type II, comprising the nonpigmented isolates, which exhibit a very broad host range.Mycobacterium avium subsp. paratuberculosis is the etiological agent of paratuberculosis (Johne's disease), a fatal chronic granulomatous enteritis affecting principally ruminants. The organism is distinguished from other subspecies of M. avium by its mycobactin requirement for in vitro growth (22) and by the presence of the insertion sequence IS900 (9). Molecular typing procedures such as pulsed-field gel electrophoresis (PFGE) and restriction fragment length polymorphism (RFLP) coupled with hybridization to IS900 (IS900-RFLP) have shown that in comparison with other pathogens, there is relatively little genetic variability within this subspecies. However, there is a small subset of isolates distinguished by their prominent yellow or orange pigment that have not been characterized fully.Pigmented M. avium subsp. paratuberculosis has been isolated in cases of lepromatous ovine paratuberculosis (18)(19)(20). In such cases, the affected intestine appears yellow or orange, probably due to the very large number of organisms present in the lesions. The organisms are difficult to culture, and few isolates have been maintained in collections. The pigmentation appears to be an inherent characteristic, as it is present at all stages of growth and is not altered by animal passage. The pigmented strains appear to have a host preference for sheep, although they can produce disease in cattle following experimental infection (21) and there has been one report of a naturally occurring bovine case (24). The purpose of this study was to analyze available pigmented isolates and investigate their genetic relatedness to other, nonpigmented M. avium subsp. paratuberculosis isolates. MATERIALS AND METHODSM. avium subsp. paratuberculosis isolates. Five pigmented and 88 nonpigmented strains of M. avium subsp. paratuberculosis were analyzed in this study. Four of the pigmented strains were isolated at the Moredun Research Institute, Penicuik, Scotland, and Finn Saxegaard (National Veterinary Institute, Oslo, Norway) kindly provided pigmented...

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Section: Resultssupporting

confidence: 63%

Section: Resultsmentioning

confidence: 86%

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Molecular Characterization of Pigmented and Nonpigmented Isolates of Mycobacterium avium subsp. paratuberculosis

et al. 2002

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“…In our study, IS900 RFLP analysis of MAP isolates, from different locations of one animal revealed that all isolates were of RFLP type C1. These results are in agreement with other studies where IS900 RFLP patterns are described as very stable over a long period of time (Cousins et al, 2000;Ayele et al, 2004). Since RFLP type C1 is the predominating pattern of MAP isolates from deer and cattle from the Czech Republic (Pavlik et al, 2000;Machackova et al, 2004;Machackova-Kopecna et al, 2005), it is not possible to conclude, without epidemiological knowledge, if MAP was transmitted from only one source of infection.…”

Section: Discussionsupporting

confidence: 79%

A mixed infection of Mycobacterium avium subsp. paratuberculosis and M. a. hominissuis in one red deer (Cervus elaphus) studied by IS900 BstEII and IS1245 PvuII RFLP analyses: a case report

Morávková¹,

Trcka²,

Lamka³

et al. 2008

Vet. Med. - Czech

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A mixed infection of (MAH) in one naturally infected red deer stag from a game park is described. The animal was euthanized because of symptoms of poor condition, weight loss and chronic diarrhoea. In spite of that, pathological lesions were observed only in the mesenteric lymph nodes, which were five to ten times enlarged with confluent caseous granulomas of 1 to 10 mm in size. Mycobacteria were isolated from all studied samples: a mixed infection of MAP and MAH was confirmed by multiplex PCR for the detection of IS900, IS901, IS1245 and dnaJ. MAP of the identical IS900 BstEII RFLP type C1 was isolated from all tissue samples and faeces. MAH isolates were detected in six examined tissue samples, including three mesenteric lymph nodes with caseous granulomas. Only minor differences in the band numbers and position of four different IS1245 PvuII RFLP patterns of MAH isolates were found. It follows from these results that red deer may potentially be infected with MAH, when a MAP infection is under way.

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“…paratuberculosis (19). Hence, IS900 has been the marker of choice for most fingerprinting studies reported (11,34,47). This exploits the fact that the IS900 elements show a high degree of target sequence specificity resulting in similar fingerprints in epidemiologically related isolates (19).…”

Section: Discussionmentioning

confidence: 99%

Molecular Epidemiology of Mycobacterium avium subsp. paratuberculosis : Evidence for Limited Strain Diversity, Strain Sharing, and Identification of Unique Targets for Diagnosis

et al. 2003

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The objectives of this study were to understand the molecular diversity of animal and human strains of Mycobacterium avium subsp. paratuberculosis isolated in the United States and to identify M. avium subsp. paratuberculosis-specific diagnostic molecular markers to aid in disease detection, prevention, and control. Multiplex PCR of IS900 integration loci (MPIL) and amplified fragment length polymorphism (AFLP) analyses were used to fingerprint M. avium subsp. paratuberculosis isolates recovered from animals (n ‫؍‬ 203) and patients with Crohn's disease (n ‫؍‬ 7) from diverse geographic localities. Six hundred bacterial cultures, including M. avium subsp. paratuberculosis (n ‫؍‬ 303), non-M. avium subsp. paratuberculosis mycobacteria (n ‫؍‬ 129), and other nonmycobacterial species (n ‫؍‬ 168), were analyzed to evaluate the specificity of two IS900 integration loci and a newly described M. avium subsp. paratuberculosis-specific sequence (locus 251) as potential targets for the diagnosis of M. avium subsp. paratuberculosis. MPIL fingerprint analysis revealed that 78% of bovine origin M. avium subsp. paratuberculosis isolates clustered together into a major node, whereas isolates from human and ovine sources showed greater genetic diversity. MPIL analysis also showed that the M. avium subsp. paratuberculosis isolates from ovine and bovine sources from the same state were more closely associated than were isolates from different geographic regions, suggesting that some of the strains are shared between these ruminant species. AFLP fingerprinting revealed a similar pattern, with most isolates from bovine sources clustering into two major nodes, while those recovered from sheep or humans were clustered on distinct branches. Overall, this study identified a high degree of genetic similarity between M. avium subsp. paratuberculosis strains recovered from cows regardless of geographic origin. Further, the results of our analyses reveal a relatively higher degree of genetic heterogeneity among M. avium subsp. paratuberculosis isolates recovered from human and ovine sources.Paratuberculosis or Johne's disease (JD) is a chronic granulomatous enteritis of ruminants caused by Mycobacterium avium subsp. paratuberculosis (48). It is estimated that 35% of the U.S. herds are infected with M. avium subsp. paratuberculosis, resulting in annual losses of $200 million (7). Crohn's disease (CD) is also a chronic inflammation of distal intestines exhibiting a pathology similar to that of JD in ruminants. The prevalence of CD is estimated to be 0.15% among the U.S. population resulting in substantial morbidity and medical costs (1). M. avium subsp. paratuberculosis has been implicated as a cause of CD in humans (13,21,36). Currently, the evidence for a link remains inconclusive since strain sharing or a causal role of M. avium subsp. paratuberculosis has not been demonstrated.Comprehensive analysis of the molecular diversity and comparative molecular pathology of M. avium subsp. paratuberculosis will help establish the degree of heterog...

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DNA fingerprinting of Australian isolates of Mycobacterium avium subsp paratuberculosis using IS900 RFLP

Cited by 54 publications

References 22 publications

Molecular Characterization of Pigmented and Nonpigmented Isolates of Mycobacterium avium subsp. paratuberculosis

Molecular Characterization of Pigmented and Nonpigmented Isolates of Mycobacterium avium subsp. paratuberculosis

A mixed infection of Mycobacterium avium subsp. paratuberculosis and M. a. hominissuis in one red deer (Cervus elaphus) studied by IS900 BstEII and IS1245 PvuII RFLP analyses: a case report

Molecular Epidemiology of Mycobacterium avium subsp. paratuberculosis : Evidence for Limited Strain Diversity, Strain Sharing, and Identification of Unique Targets for Diagnosis

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