A rapid method for the extraction and detection of Mycobacterium avium subspecies paratuberculosis from clinical specimens

Challans, JA; Stevenson, Karen; Reid, HW; Sharp, JM

doi:10.1136/vr.134.4.95

Cited by 38 publications

(31 citation statements)

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“…The assay is specific for M. paratuberculosis [11][12][13], but it has been reported to have a poor sensitivity when used on clinical specimens such as faeces and tissues because of unknown inhibitors and difficulty in sample preparation [11,23,26]. In our hands the limit of detection was 10-100 organisms per gram of tissue.…”

Section: Discussionmentioning

confidence: 65%

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An abattoir-based study of the prevalence of subclinical Johne's disease in adult cattle in south west England

1996

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SUMMARYThe prevalance of subclinical Johne's disease was estimated in adult cattle slaughtered at three major abattoirs in south west England. A polymerase chain reaction (PCR) based on IS900 was used to detect Mycobacterium paratuberculosis in intestinal lymph nodes of 1553 cattle. Culture was also carried out on all PCR positive and inconclusive samples. The prevalence of subclinical disease in adult cattle was 3 5 % (95 % confidence intervals (CI) 2 6-47) by PCR and 2 6 % (CI 1-8-3 6) by culture. The proportion of the disease in each month ranged from 1-6 % (CI 02-5 5) in April to 4-6 % (CI 2 8-6 9) in November, but the difference was not significant (P > 005). The proportion of PCR positive lymph nodes in each abattoir ranged from 2-8 % (CI 1 6-4-6) to 4 9 % (CI 2 9-7 6), this difference was not significant either (P > 005). The prevalence in young cattle was 2-0 % (CI 0 6-4A5). The difference between age groups was not statistically significant (P > 005).

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Section: Discussionmentioning

confidence: 65%

“…Sample preparation A modification of the method reported by Challans and colleagues [23] was used for DNA extraction. Briefly, 1 g of sample was mixed with 5 ml distilled water and mashed up in a stomacher (Seward Medical, model 80) for 2 min at high speed.…”

Section: Methodsmentioning

confidence: 99%

An abattoir-based study of the prevalence of subclinical Johne's disease in adult cattle in south west England

1996

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“…The specificity of the insertion sequence IS900 has been demonstrated by different investigators (Kunze et al, 1992;Challans et al, 1994;Pe´rez et al, 1994;Miller et al, 1999;Coetsier et al, 2000;Garrido et al, 2000;Collins et al, 2002) and may provide the basis for an alternative to culture. The technique failed, however, to detect a significant proportion of animals with focal lesions; this may have been due to inability to detect small numbers of mycobacteria or to the presence of unusual forms of mycobacteria (Pe´rez et al, 1994).…”

Section: Discussionmentioning

confidence: 99%

“…The PCR technique as described by Challans et al (1994) was used to examine fresh tissue samples from the jejunum, ileum, ileocaecal valve and associated lymph nodes. RJ1-PT91 primers were selected to amplify a 388 bp fragment of the IS900 insertion sequence specific for Map.…”

Section: Polymerase Chain Reaction (Pcr)mentioning

confidence: 99%

Histopathological Classification of Lesions Observed in Natural Cases of Paratuberculosis in Free-ranging Fallow Deer (Dama dama)

Balseiro

Marı́n

Solano

et al. 2008

Journal of Comparative Pathology

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SummaryNinety-five adult fallow deer, legally hunted in the Regional Hunting Reserve of El Sueve (Northern Spain), were subjected to a post-mortem examination for paratuberculosis, samples being taken from the proximal and distal jejunum, proximal and distal ileum, ileocaecal valve and associated lymph nodes. The lesions were divided into four categories. Focal lesions (n ¼ 19 cases) consisted of small granulomas, mainly in the jejunal and ileal lymph nodes. Multifocal lesions (n ¼ 4) consisted of well-demarcated granulomas in the intestinal lymphoid tissue and also in the intestinal lamina propria. Diffuse multibacillary lesions (n ¼ 2) were characterized by a severe granulomatous enteritis and lymphadenitis. Macrophages and numerous Langhans giant cells containing many mycobacteria were present, resulting in macroscopical changes in the normal gut morphology. These changes were found from the proximal jejunum to the ileocaecal valve, but lesions were always particularly severe in the distal jejunum. In diffuse intermediate (multibacillaryelymphocytic) lesions (n ¼ 3) the infiltrate consisted of lymphocytes, macrophages and Langhans giant cells, with small numbers of mycobacteria. Mycobacterium avium subspecies paratuberculosis was identified by a polymerase chain reaction technique. The widespread occurrence of paratuberculosis in fallow deer in this Reserve represents a potential source of infection for other susceptible species.

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“…The cells were transferred to a screw-cap microfuge tube containing 1 ml washed 0n1 mm zirconium\silica beads. Cells were disrupted by bead-beating in a Mini Bead-Beater (BioSpec Products) (Challans et al, 1994). Five beating cycles of 1 min each were separated by 1 min cooling intervals on ice to avoid overheating and possible denaturation of the antigen preparation.…”

Section: Methodsmentioning

confidence: 99%

Unique expression of a highly conserved mycobacterial gene in IS901 + Mycobacterium avium The GenBank accession number for the p40 gene, together with 542 bp upstream sequence, is AF247653.

et al. 2001

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A rapid method for the extraction and detection of Mycobacterium avium subspecies paratuberculosis from clinical specimens

Cited by 38 publications

References 0 publications

An abattoir-based study of the prevalence of subclinical Johne's disease in adult cattle in south west England

An abattoir-based study of the prevalence of subclinical Johne's disease in adult cattle in south west England

Histopathological Classification of Lesions Observed in Natural Cases of Paratuberculosis in Free-ranging Fallow Deer (Dama dama)

Unique expression of a highly conserved mycobacterial gene in IS901 + Mycobacterium avium The GenBank accession number for the p40 gene, together with 542 bp upstream sequence, is AF247653.

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