Development and evaluation of a gamma-interferon assay for tuberculosis in badgers (Meles meles)

Deanna Dalley; Dipesh Davé; Sandrine Lesellier; Simonette Palmer; Timothy Crawshaw; R. Glyn Hewinson; Mark Chambers

doi:10.1016/j.tube.2007.11.001

Development and evaluation of a gamma-interferon assay for tuberculosis in badgers (Meles meles)

Deanna Dalley, Dipesh Davé, Sandrine Lesellier, Simonette Palmer, Timothy Crawshaw, R. Glyn Hewinson, Mark Chambers

Animal and Plant Health Agency

Research output: Contribution to journal › Article › peer-review

67 Citations (Scopus)

Abstract

In this paper we report the development of a sensitive and specific assay for the detection of tuberculosis (TB) in European badgers (Meles meles), based on the stimulation of lymphocytes in whole-blood culture and the subsequent detection of gamma-interferon (IFNγ) by sandwich ELISA. The comparative levels of IFNγ produced to bovine and avian tuberculin (B-A) was used as the basis of determining the TB status of badgers, resulting in a more sensitive test than that based on the defined Mycobacterium bovis antigens ESAT6 and CFP10. The assay was evaluated using 235 badgers. The IFNγ EIA (enzyme immunoassay) based on a monoclonal pair (mEIA) was more sensitive than one using a rabbit polyclonal antiserum (pEIA). At a specificity of 93.6%, the mEIA was 80.9% sensitive, compared to a sensitivity of 74.5% for the pEIA. At the same specificity as the EIA, the current serological ELISA test for TB in badgers (Brock test) had a sensitivity of 48.9%. Only one of the culture positive badgers missed by the mEIA was correctly diagnosed by the Brock test, suggesting that the combination of both a T-cell and serological test has little diagnostic advantage. Crown

Original language	English
Pages (from-to)	235-243
Number of pages	9
Journal	Tuberculosis
Volume	88
Issue number	3
DOIs	https://doi.org/10.1016/j.tube.2007.11.001
Publication status	Published - 01 May 2008

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10.1016/j.tube.2007.11.001

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title = "Development and evaluation of a gamma-interferon assay for tuberculosis in badgers (Meles meles)",

abstract = "In this paper we report the development of a sensitive and specific assay for the detection of tuberculosis (TB) in European badgers (Meles meles), based on the stimulation of lymphocytes in whole-blood culture and the subsequent detection of gamma-interferon (IFNγ) by sandwich ELISA. The comparative levels of IFNγ produced to bovine and avian tuberculin (B-A) was used as the basis of determining the TB status of badgers, resulting in a more sensitive test than that based on the defined Mycobacterium bovis antigens ESAT6 and CFP10. The assay was evaluated using 235 badgers. The IFNγ EIA (enzyme immunoassay) based on a monoclonal pair (mEIA) was more sensitive than one using a rabbit polyclonal antiserum (pEIA). At a specificity of 93.6%, the mEIA was 80.9% sensitive, compared to a sensitivity of 74.5% for the pEIA. At the same specificity as the EIA, the current serological ELISA test for TB in badgers (Brock test) had a sensitivity of 48.9%. Only one of the culture positive badgers missed by the mEIA was correctly diagnosed by the Brock test, suggesting that the combination of both a T-cell and serological test has little diagnostic advantage. Crown",

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AU - Crawshaw, Timothy

AU - Hewinson, R. Glyn

AU - Chambers, Mark

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AB - In this paper we report the development of a sensitive and specific assay for the detection of tuberculosis (TB) in European badgers (Meles meles), based on the stimulation of lymphocytes in whole-blood culture and the subsequent detection of gamma-interferon (IFNγ) by sandwich ELISA. The comparative levels of IFNγ produced to bovine and avian tuberculin (B-A) was used as the basis of determining the TB status of badgers, resulting in a more sensitive test than that based on the defined Mycobacterium bovis antigens ESAT6 and CFP10. The assay was evaluated using 235 badgers. The IFNγ EIA (enzyme immunoassay) based on a monoclonal pair (mEIA) was more sensitive than one using a rabbit polyclonal antiserum (pEIA). At a specificity of 93.6%, the mEIA was 80.9% sensitive, compared to a sensitivity of 74.5% for the pEIA. At the same specificity as the EIA, the current serological ELISA test for TB in badgers (Brock test) had a sensitivity of 48.9%. Only one of the culture positive badgers missed by the mEIA was correctly diagnosed by the Brock test, suggesting that the combination of both a T-cell and serological test has little diagnostic advantage. Crown

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KW - Interferon gamma

KW - Mycobacterium bovis

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Development and evaluation of a gamma-interferon assay for tuberculosis in badgers (Meles meles)

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