A reverse transcriptase (RT)-nested-polymerase chain reaction (PCR) was standardised to detect bovine respiratory syncytial virus (BRSV), using a Brazilian isolate, in three experimentally infected calves. This followed initial tests in infected chicken embryo related (CER) cells. One animal had lesions, characterized by interstitial multifocal pneumonia, severe interstitial and subpleural emphysema, and lung consolidated areas. Lung and tracheal tissues collected 6 days after infection were analysed by RT-nested-PCR. Primers, specific for the BRSV G and F glycoproteins genes, yielded amplification fragments of 371 and 481 bp, respectively, from the RNA of the cell-propagated virus. Using RNA extracted from organs of infected calves, RT-nested-PCR amplified the fragment of the G gene in all tracheal samples, but in only two of three lung samples analysed. These results suggest that RT-nested-PCR could be a promising assay for diagnosis and epidemiological analysis of BRSV in Brazil.105131-5