A lymphocyte transformation assay for the detection of Mycobacterium bovis infection in the Eurasian Badger (Meles meles)

Dalley, Deanna; Chambers, Mark A.; Cockle, P; Pressling, W; Gavier‐Widén, Dolores; Hewinson, R. Glyn

doi:10.1016/s0165-2427(99)00072-0

Cited by 36 publications

(15 citation statements)

References 27 publications

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“…Acid-fast bacteria could not be detected in cytospin smears following transtracheal lavage of both animals. The lymphocyte transformation assay (LTA) for TB [9] was carried out on one ill animal (E) and found to be positive. Animal D died suddenly on November 21, 2004.…”

Section: Investigation: University Veterinary Hospitalmentioning

confidence: 99%

Tuberculosis in alpaca (Lama pacos) on a farm in Ireland. 1. A clinical report

Ryan

Dwyer²,

Connolly³

et al. 2008

Ir Vet J

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This case report describes tuberculosis (TB) due to infection with Mycobacterium bovis (M. bovis) in alpaca (Lama pacos) on a farm in Ireland. Two severely debilitated alpaca were presented to the University Veterinary Hospital, University College Dublin in November 2004. Bloods were taken, and haematology and biochemistry results were indicative of chronic infection. Radiological examination showed evidence of diffuse granulomatous pneumonia suggestive of tuberculosis. On necropsy there were granulomatous lesions present throughout many body organs including lung, liver, kidney, intestine as well on peritoneum and mesentery. Culture of acid-fast bacilli from lesions led to a diagnosis of tuberculosis due to M. bovis. The use of intradermal skin testing proved inefficient and unreliable for ante mortem diagnosis of tuberculosis in alpaca. Infection due to M. bovis should be considered among the differential diagnoses of debilitating diseases in alpaca, particularly those farmed in areas known to be traditional black spots for tuberculosis in cattle.

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Section: Investigation: University Veterinary Hospitalmentioning

confidence: 99%

Tuberculosis in alpaca (Lama pacos) on a farm in Ireland. 1. A clinical report

Ryan

Dwyer²,

Connolly³

et al. 2008

Ir Vet J

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“…Antibody-based assays have been developed (13,14), but they lack sensitivity (3,14), as they do for cattle (23). In comparison, assays based on measurement of cellular responses produce better results (6,23). However, those developed to date for badgers are impractical for routine use (6).…”

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confidence: 99%

“…In comparison, assays based on measurement of cellular responses produce better results (6,23). However, those developed to date for badgers are impractical for routine use (6). Tests analogous to that developed for the diagnosis of TB in cattle by the detection of gamma interferon (IFN-␥) in whole-blood cell cultures (21) have yet to be validated for badgers, although the badger IFN-␥ gene has been cloned and a polyclonal antiserum to the cytokine has been produced (7).…”

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confidence: 99%

Development and Evaluation of a Test for Tuberculosis in Live European Badgers (Meles meles) Based on Measurement of Gamma Interferon mRNA by Real-Time PCR

et al. 2007

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A real-time PCR assay for the measurement of gamma interferon (IFN-␥) mRNA in European badger (Meles meles) blood cultures was developed. The levels of IFN-␥ mRNA in blood cultures stimulated with either bovine or avian tuberculin or specific mycobacterial antigens were compared with those in a nonstimulated control blood culture as the basis for determining the tuberculosis (TB) status of live badgers. The assay was validated by testing 247 animals for which there were matching data from postmortem examination and culture of tissues. Relative changes in the levels of IFN-␥ mRNA in response to bovine tuberculin and specific antigens were found to be greater among badgers with tissues positive for TB on culture. The test was at its most accurate (87% of test results were correct) by using blood cultures containing bovine tuberculin as the antigen and when the response to avian tuberculin was taken into account by subtracting the avian tuberculin response from the bovine tuberculin response. At a specificity of 90.7%, the test was 70.6% sensitive. At the same specificity, the current serological enzyme-linked immunosorbent assay for TB in badgers was only 53% sensitive. This work demonstrates that measurement of IFN-␥ mRNA by real-time PCR is a valid method for the detection of TB in live badgers and may provide an alternative to the current serological methods of diagnosis, the Brock test. The testing procedure can be completed within 5 h of receipt of the blood culture samples. In addition, the use of a molecular biology-based test offers the potential to fully automate the testing procedure through the use of robotics.There has been a sharp rise in the incidence of bovine tuberculosis (TB) in cattle in the United Kingdom since the early 1990s (1). This adversely affects animal health and welfare and is a cause of considerable economic loss to farmers and the government. The European badger (Meles meles) has been implicated in the spread of disease by acting as a source of Mycobacterium bovis infection in cattle (9, 10). The accurate diagnosis of M. bovis infection in badgers can be achieved by bacteriological culture, but only for samples obtained postmortem. In vitro diagnostics that can be used to test live animals are required, for instance, to further develop vaccination strategies. Antibody-based assays have been developed (13, 14), but they lack sensitivity (3, 14), as they do for cattle (23). In comparison, assays based on measurement of cellular responses produce better results (6, 23). However, those developed to date for badgers are impractical for routine use (6). Tests analogous to that developed for the diagnosis of TB in cattle by the detection of gamma interferon (IFN-␥) in whole-blood cell cultures (21) have yet to be validated for badgers, although the badger IFN-␥ gene has been cloned and a polyclonal antiserum to the cytokine has been produced (7). Having cloned the badger IFN-␥ gene, we investigated the possibility of using IFN-␥ mRNA levels as a measure of the cell-mediated response to myco...

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“…Diagnosis of bTB in live badgers has been demonstrated using assays of both serological (4,20) and cell-mediated (8,9) immunity. While isolation of M. bovis from clinical samples is definitive, it is too insensitive for badgers, as infected animals yield positive samples infrequently and intermittently (3).…”

mentioning

confidence: 99%

Evaluation of a Rapid Serological Test for the Determination of Mycobacterium bovis Infection in Badgers ( Meles meles ) Found Dead

Chambers¹,

Lyashchenko²,

Greenwald³

et al. 2010

Clin Vaccine Immunol

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Mycobacterium bovis infection is the cause of bovine tuberculosis (bTB) in a wide range of mammal species, including domestic livestock and captive and free-ranging wildlife. Bovine TB remains an important zoonotic disease with significant impacts on the economy in many countries (6,22,23). Eurasian badgers (Meles meles) are a wildlife maintenance host of bTB in Great Britain and Ireland (5,15) and are implicated in the maintenance and onward transmission of M. bovis infection to cattle (10, 19).Surveillance of wildlife vectors of disease for prevalence estimates of infection may be valuable in disease control strategies and for the assessment of risk of transmission to livestock. Diagnosis of bTB in live badgers has been demonstrated using assays of both serological (4, 20) and cell-mediated (8, 9) immunity. While isolation of M. bovis from clinical samples is definitive, it is too insensitive for badgers, as infected animals yield positive samples infrequently and intermittently (3). A rapid serological test (BrockTB Stat-Pak; Chembio Diagnostic Systems, Inc.) has recently been developed for the diagnosis of bTB in multiple wildlife species (20). The test has modest sensitivity (46 to 55%) for antibody detection in live, infected badgers, but it has the advantages of being simple, rapid, inexpensive, and suitable for field application. Its utility as an animal-side test for badgers, however, is limited by the difficulties associated with obtaining a blood sample from a nonanesthetized animal.Where carcasses are recovered and submitted for mycobacterial culture, the sensitivity of diagnosis depends on the effort taken for careful examination and on the number of tissue samples submitted for culture testing and histopathology (7), as well as on the condition of the carcass. In many cases, the cost involved may prove prohibitive. Reliance on the presence of visible lesions as indicative of bTB is fraught with difficulties, as infected animals may present with no visible lesions or lesions may be the result of other infections while having the appearance of bTB (reviewed in reference 13). The purpose of this study was to determine whether the BrockTB Stat-Pak test could detect M. bovis antibody in blood collected from the carcasses of dead badgers as an alternative means of diagnosis and decision making. Animals were obtained as part of a separate government-funded study to determine the prevalence of bTB in badgers found dead in Wales (http://new.wales.gov.uk/depc/ publications/environmentandcountryside/animalhealthandwelfare/ diseasesurveillancecontrol/bovinetb/2567889/publicationindex/ 2326585/badgerfounddeadreport?langϭen). Our results reveal that the BrockTB Stat-Pak test used on thoracic blood samples

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A lymphocyte transformation assay for the detection of Mycobacterium bovis infection in the Eurasian Badger (Meles meles)

Cited by 36 publications

References 27 publications

Tuberculosis in alpaca (Lama pacos) on a farm in Ireland. 1. A clinical report

Tuberculosis in alpaca (Lama pacos) on a farm in Ireland. 1. A clinical report

Development and Evaluation of a Test for Tuberculosis in Live European Badgers (Meles meles) Based on Measurement of Gamma Interferon mRNA by Real-Time PCR

Evaluation of a Rapid Serological Test for the Determination of Mycobacterium bovis Infection in Badgers ( Meles meles ) Found Dead

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