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Background  The European badger is an important wildlife reservoir of  Mycobacterium bovis  implicated in the spread of bovine tuberculosis in the United Kingdom and Ireland. Infected badgers are known to shed  M. bovis  in their urine and faeces, which may contaminate the environment. To aid bovine tuberculosis control efforts novel diagnostic tests for detecting infected and shedding badgers are needed. We proposed development of a novel, rapid immunochromatographic lateral flow device (LFD) as a non-invasive test to detect  M. bovis  cells in badger faeces. Its application in combination with immunomagnetic separation (IMS) to detect  Mycobacterium bovis  cells in badger faeces is reported here.    Results  A novel prototype LFD for  M. bovis  cells was successfully developed, with unique specificity for  M. bovis  and a limit of detection 50% (LOD 50% ) of 1.7 × 10 4   M. bovis  cells/ml. When IMS was employed to selectively capture and concentrate  M. bovis  cells from badger faeces prior to LFD testing, the LOD 50%  of the IMS-LFD assay was 2.8 × 10 5   M. bovis  cells/ml faecal homogenate. Faeces samples collected from latrines at badger setts in a region of endemic bovine tuberculosis infection were tested; 78 (18%) of 441 samples tested IMS-LFD assay positive, whereas 140 (32%) tested IMS-qPCR positive (Kappa agreement −0.009 ± 0.044,  p  = 0.838). Subsequently, when 130 faeces samples from live captured, or captive, badgers of known infection status (on the basis of StatPak, interferon-γ and/or culture results) were tested, the IMS-LFD assay had higher relative diagnostic specificity (Sp 0.926), but poorer relative diagnostic sensitivity (Se 0.081), than IMS-qPCR (Sp 0.706, Se 0.581) and IMS-culture (Sp 0.794, Se 0.436).    Conclusions  The novel IMS-LFD assay, although very specific for  M. bovis , has low analytical sensitivity (indicated by the LOD 50% ) and would only detect badgers shedding high numbers of  M. bovis  (>10 4–5  cells/g) in their faeces. The novel LFD would, therefore, have limited value as a non-invasive test for badger TB surveillance purposes but it may have value for alternative veterinary diagnostic applications.

bmc veterinary research

Development of a novel immunochromatographic lateral flow assay specific for Mycobacterium bovis cells and its application in combination with immunomagnetic separation to test badger faeces