Development of oligonucleotide microarray system for differential diagnosis of enteric viruses in diarrheic fecal samples in pigs
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Abstract
An oligonucleotide microarray system was developed for the simultaneous detection of porcine epidemic diarrhea virus, transmissible gastroenteritis virus, porcine enteric calicivirus, porcine group A and C rotavirus. RNAs of the reference viruses and porcine diarrhea samples were extracted and amplified using onestep multiplex RTPCR in the presence of cyanine 5dCTP and hybridized on the microarray chip that spotted the virusspecific oligonucleotides. This system were approximately 10 to 100fold higher in sensitivity than conventional RTPCR, and the assay time was less than 3 hours. The relative sensitivity and specificity were 92% and 72.2%, respectively, based on 102 porcine diarrhea samples using RTPCR as gold standard. These results suggested that the oligonucleotide microarray system in this study be probably more reliable and reproducible means for detecting porcine enteric viruses and that it could be of substantial use in routine diagnostic laboratories.
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