Reverse Transcription-pcr for Detection of Porcine Diarrhea Associated Group a Rotavirus in Field Samples from Porcines Farmed in North-eastern Romania

Group A rotaviruses (GARVs) cause acute diarrhea and malabsorption in new-born and young piglets, resulting in high mortality and morbidity. Evidence of this infection has been reported in various European countries. However, there is little evidence of porcine GARV infections in Romania. The aim of this study was the detection of the GARV in an outbreak of diarrhea in piglets and sows farmed in North-Eastern Romania. We examined 25 fecal samples: 20 diarrheic fecal specimens collected from piglets and 5 normal fecal samples collected from healthy sows raised in a closed-circuit farm. The extracted ARN underwent a reverse transcription step, followed by a classical polymerase chain reaction assay, using primers able to amplify a fragment of 317 bp from NSP5 gene, the most conservative gene among GARV strains and isolates. RT-PCR using specific primer for the GARV NSP5 gene detected GARV-positive reactions in 15 (60%) fecal samples. Of these, 12 out of 20 diarrheic fecal samples (60%) and 3 out of 5 fecal samples (60%) tested positive for porcine GARVs. The data showed that GARV was identified in the vast majority of both diarrheic and normal fecal samples, suggesting that the GARV may represent a major pathogen with an important role in this diarrhea outbreak. Thus, this RT-PCR assay proved to be a rapid and precise diagnosis assay for detection of porcine GARV. Furthermore, the primers annealing temperature (60 C) is able to confer to this assay an increased specificity and sensitivity. In order to prevent the economical loses, the use of a reliable diagnosis method allowing the detection of rotavirus could contribute in achieving this goal, together with the identification and removal of the asymptomatic carriers.