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Background  Culture of  M. bovis  from diagnostic specimens is the gold standard for bovine tuberculosis diagnostics in the USA. Detection of  M. bovis  by PCR in tissue homogenates may provide a simple rapid method to complement bacterial culture. A significant impediment to PCR based assays on tissue homogenates is specificity since mycobacteria other than  M. bovis  may be associated with the tissues.    Results  Previously published IS 6110  based PCR diagnostic assays, along with one developed in house, were tested against environmental mycobacteria commonly isolated from diagnostic tissues submitted to the National Veterinary Services Laboratory. A real-time PCR assay was developed (IS6110_T) that had increased specificity over other IS 6110  based assays. Of the 13 non-tuberculous mycobacteria tested with IS6110_T only  M. wolinskyi  was positive. Thirty  M. bovis  infected tissue homogenates and 18 control tissues were used to evaluate the potential for the assay as a diagnostic test. In this small sample, IS6110_T detected 20/30 samples from  M. bovis  infected animals and 0/18 control tissues.    Conclusions  The IS6110_T assay provides a PCR based assay system that is compatible with current diagnostic protocols for the detection of  M. bovis  in the USA and compliments current testing strategies.

Improved specificity for detection of Mycobacterium bovis in fresh tissues using IS6110 real-time PCR