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KMID : 1023520070300040489
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Korean Journal of Veterinary Service
2007 Volume.30 No. 4 p.489 ~ p.496
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Development of oligonucleotide microarray system for differential diagnosis of enteric viruses in diarrheic fecal samples in pigs
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Park Nam-Yong
Kim Yong-Hwan
Cho Ho-Seong
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Abstract
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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 one-step multiplex RT-PCR in the presence of cyanine 5-dCTP and hybridized on the microarray chip that spotted the virus-specific oligonucleotides. This system were approximately 10-to 100-fold higher in sensitivity than conventional RT-PCR, 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 RT-PCR 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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KEYWORD
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Microarray, porcine enteric viruses, DNA chip
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