Group D rotaviruses (RVs-D) have been documented in birds and, while they may be common in these
animals, few molecular studies are available for this specific group. In this study, specific primers for
the gene that encodes for the RVs-D VP6 protein were designed and used in a reverse transcription
polymerase chain reaction (RT-PCR). Thirty pools of samples were tested by polyacrylamide gel electrophoresis
(PAGE) yielding a 30% (9/30) positivity. These pools were subjected subsequently to RT-PCR,
with a 53% (16/30) positivity rate. The sensitivity of the PCR assay was demonstrated up to a dilution
of 5 × 10−4 ng/L (0.5 pg/L) of the cloned VP6 gene. The four samples were sequenced and showed
90.8–91.1% similarity with regards to the RVs-D VP6 gene. To assess for specificity our RT-PCR was applied
to nine samples known to contain enteric viral agents other than group D rotaviruses including picobirnavirus,
rotavirus group A, and reovirus with negative results. Overall, the data confirm the specificity of
the primers used for detecting the RVs-D by RT-PCR, suggesting that this assay can be used for diagnostic
purposes.