P08

Bovine tuberculosis is a chronic granulomatous disease mainly affecting lymph node and lung tissues of cattle. Lesions found at post-mortem examinations can be confirmed by bacterial isolation of Mycobacterium bovis. Thus is laborious and time consuming and may require months, which slows the control and eradication of the disease. The objective was to demonstrate the usefulness of real-time PCR and RD4-PCR as molecular methods for rapid detection of bTB in Bulgaria. The study panel included 68 tissue samples (lymph nodes) from cattle from 3 farms in 2 regions of Bulgaria. Culture tests and histology of lymph nodes both with and without visible lesions are undertaken to isolate and type the causative strain. The DNA was isolated with DNeasy blood and tissue kit (Qiagen). RT-PCR was provided by Bio-T kit® MTBC (Biosellal). RD4-PCR was used to differentiate the M. bovis from M. caprae. By RT- PCR, the affiliation to the MTВ complex was demonstrated for all 68 studied samples. RD4-PCR subdivided the samples into 62 M.caprae and 6 M.bovis strains. The observed identification of M. caprae  can be indicative of a geographical-type appearance in Bulgaria. The application of RT- PCR is convenient method for rapid detection of M. bovis from cattle in the Bulgarian settings. RD4-PCR provided a sufficiently high differentiation and may be used for a first-line typing. This proves the need for the simultaneous application of both methods in the diagnosis of bovine tuberculosis in Bulgaria. This work was supported by the grant DN16/12, National Science Fund, Bulgaria.