Use of BACTEC radiometric culture method and polymerase chain reaction for the rapid screening of faeces and tissues for Mycobacterium paratuberculosis

Cousins, DV; Evans, R. J.; Francis, B.R.

doi:10.1111/j.1751-0813.1995.tb03489.x

Cited by 51 publications

(48 citation statements)

References 8 publications

(2 reference statements)

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“…The growth of the two bovine isolates identified as M. avium subsp. paratuberculosis depended on the presence of mycobactin in the medium and they were positive in a PCR test using specific primers to amplify the insertion sequence IS900, as described by Cousins et al (1995). The identification for reference strains including M. scrofulaceum has been established previously by Wayne et al (1993).…”

Section: Bacterial Isolates and Their Identification A Total Of 97mentioning

confidence: 99%

Use of multilocus enzyme electrophoresis to examine genetic relationships amongst isolates of Mycobacterium intracellulare and related species

Feizabadi

Robertson

Cousins³

et al. 1997

Microbiology

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As part of a larger study investigating diversity and distribution of Mycobacterium spp. in Australia, multilocus enzyme electrophoresis was used to assess genetic relationships a t 17 enzyme loci amongst a collection of reference strains and isolates initially identified on biochemical and other grounds as M. intracellulare (70), 'X' mycobacteria (lo), M. scrofulaceum (7), M. avium (8) and M. avium subsp. paratuberculosis (2). Two of the isolates initially identified as M. intracellulare were shown to be quite distinct from the others. Both gave negative results in a species-specific DNA probe test, whilst one was positive by PCR. These results emphasize the uncertainties involved in identifying members of this group. The other M. intracellulare isolates formed a cohesive but diverse group, being divided into 48 electrophoretic types (ETs), with a mean genetic diversity of 0.38. Forty-three of these ETs contained only single isolates. There was no clear relationship between the serovar and E l designation. The index of association calculated for M. intracellulare was significantly different from zero, suggesting that it is a clonal species. PFGE was also applied to selected isolates from the ETs containing multiple isolates, and some of these could be differentiated further. The strains of M. scrofulaceum and 'X' mycobacteria were distinct from M. intracellulare, but themselves were highly heterogeneous, with mean genetic diversities of 0.66 and 0-65, respectively. Each of these groups may represent more than one species. M. avium strains were distinct from the two M. avium subsp. paratuberculosis strains, as well as from the other mycobacteria studied.

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Section: Bacterial Isolates and Their Identification A Total Of 97mentioning

confidence: 99%

Use of multilocus enzyme electrophoresis to examine genetic relationships amongst isolates of Mycobacterium intracellulare and related species

Feizabadi

Robertson

Cousins³

et al. 1997

Microbiology

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“…Culture positive samples were further confirmed by polymerase chain reaction and restriction endonuclease analysis by demonstrating the presence of IS900 (Cousins et al, 1995;Whittington et al, 1998). The OJD prevalence of each cohort was calculated from PFC results using a Bayesian model for variable pool size (Dhand et al, 2010).…”

Section: Pooled Faecal Culture (Pfc)mentioning

confidence: 98%

Comparison of pre- and post-vaccination ovine Johne's disease prevalence using a Bayesian approach

Dhand

Johnson

Eppleston

et al. 2013

Preventive Veterinary Medicine

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This study was conducted to evaluate the effectiveness of Gudair TM vaccine in decreasing the prevalence of shedding of Mycobacterium avium subsp. paratuberculosis (MAP) in flocks of varying initial prevalence. Thirty seven self-replacing Merino flocks from New South Wales and Victoria (Australia) that had been vaccinating lambs with Gudair TM for at least five years were enrolled in the study. These flocks had been tested prior to or at commencement of vaccination using pooled faecal culture, agar gel immunodiffusion or both tests. These prevaccination test results were used to estimate pre-vaccination prevalence. Post-vaccination prevalence was estimated from culture of usually 7 pools of 50 sheep collected from the enrolled flocks in [2008][2009], approximately five or more years after commencement of vaccination.A Bayesian model was developed to estimate and compare the pre-and postvaccination prevalences for the enrolled flocks. Apparent pre-and post-vaccination prevalences for flocks were modelled as functions of the true pre-and post-vaccination prevalences, respectively, and the sensitivities and specificities of the respective diagnostic tests. Logit-normal models were specified on pre-and post-vaccination true prevalences and were then used to make inferences about the median and 90 th percentile of the prevalence 2 distributions and their differences. Priors were mostly specified based on published literature or analysis of abattoir surveillance data for this population of flocks.The analysis found a significant decline in ovine Johne's disease prevalence from a pre-vaccination median prevalence of 2.72% [95% probability interval (PI): 1.40; 6.86%] to a post-vaccination median prevalence of 0.72% (0.39; 1.27%). However 30 of the 37 flocks still contained sheep that were shedding MAP in their faeces. The results suggest that vaccination with Gudair™ is usually effective in reducing the prevalence of faecal shedding but the response to vaccination is variable among flocks. This approach could be implemented in similar situations to compare prevalences where information from multiple diagnostic tests with varied sensitivities and specificities is available.

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“…It can detect a pathogen by multiple gene targets simultaneously, as an alternative to successive corroboration of positive samples. Cousins et al (1995) and Tasara et al (2005) used multiplex PCR for MAP detection in conventional PCR setups which can be combined with real-time PCR Slana et al 2008). However, it makes its optimization complex and shows lower sensitivity due to reagents interference and primer dimer formation (Rachlin et al 2005).…”

Section: Radioisotopic Culturementioning

confidence: 99%

Trends and advances in the diagnosis and control of paratuberculosis in domestic livestock

Chaubey

Gupta

et al. 2016

Veterinary Quarterly

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Paratuberculosis (pTB) is a chronic granulomatous enteritis caused by Mycobacterium avium subsp. paratuberculosis (MAP) in a wide variety of domestic and wild animals. Control of pTB is difficult due to the lack of sensitive, efficacious and cost-effective diagnostics and marker vaccines. Microscopy, culture, and PCR have been used for the screening of MAP infection in animals for quite a long time. Besides, giving variable sensitivity and specificity, these tests have not been considered ideal for large-scale screening of domestic livestock. Serological tests like ELISA easily detects anti-MAP antibodies. However, it cannot differentiate between the vaccinated and infected animals. Nanotechnology-based diagnostic tests are underway to improve the sensitivity and specificity. Newer generation diagnostic tests based on recombinant MAP secretory proteins would open new paradigm for the differentiation between infected and vaccinated animals and for early detection of the infection. Due to higher seroreactivity of secretory proteins vis-a-vis cellular proteins, the secretory proteins may be used as marker vaccine, which may aid in the control of pTB infection in animals. Secretory proteins can be potentially used to develop future diagnostics, surveillance and monitoring of the disease progression in animals and the marker vaccine for the control and eradication of pTB.

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Use of BACTEC radiometric culture method and polymerase chain reaction for the rapid screening of faeces and tissues for Mycobacterium paratuberculosis

Cited by 51 publications

References 8 publications

Use of multilocus enzyme electrophoresis to examine genetic relationships amongst isolates of Mycobacterium intracellulare and related species

Use of multilocus enzyme electrophoresis to examine genetic relationships amongst isolates of Mycobacterium intracellulare and related species

Comparison of pre- and post-vaccination ovine Johne's disease prevalence using a Bayesian approach

Trends and advances in the diagnosis and control of paratuberculosis in domestic livestock

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