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Development of oligonucleotide microarray system for differential diagnosis of enteric viruses in diarrheic fecal samples in pigs  

Park, Nam-Yong (Department of Veterinary Pathology, College of Veterinary Medicine, Chonnam National University)
Kim, Yong-Hwan (Department of Veterinary Research, Gwangju City Institute of Health and Environment)
Cho, Ho-Seong (Department of Veterinary Pathology, College of Veterinary Medicine, Chonnam National University)
Publication Information
Korean Journal of Veterinary Service / v.30, no.4, 2007 , pp. 489-496 More about this Journal
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 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.
Keywords
Microarray; porcine enteric viruses; DNA chip;
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1 Call DR, Brockman FJ, Chandler DP. 2001, Detecting and genotyping Escherichia coli O157:H7using multiplexed PCR and nucleic acid microarrays. Int J Food Microbial 67 (1-2) :71-80   DOI   ScienceOn
2 Murphy FA, Gibb E, Horzinek MC, et al. 1999, Laboratory diagnosis of viral diseases. In Veterinary virology, 3 Eds. Academic press, Calif: 193-224
3 Rozen S, Skaletsky H. 2000, Primer3 on the WWW for general users and for biologist programmers. Methods Mol Biol 132:365-386
4 Straw BE, Dewey CE, Wilson MR. 2006, Differential diagnosis of swine diseases. In Diseases of swine, 9 Eds. Iowa State University Press, Ames, IA: 241-283
5 Wang RF, Beggs ML, Robertson LH, et al. 2002, Design and evaluation of oligonucleotide-microarray method for the detection of human intestinal bacteria in fecal samples. FEMS Microbiol Lett 123(2):175-182
6 Elschner M, Prudlo J, Hotzel H, et al. 2002, Nested reverse transcriptasepolymerase chain reaction for the detection of group A rotaviruses. J Vet Med B 49 (2) :77-81   DOI   ScienceOn
7 Chizhikov V, Rasooly A, Chumakov K, et al. 2001, Microarray analysis of microbial virulence factors. Appl Environ Microbiol 67 (7) :3258-3263   DOI   ScienceOn
8 Neverov AA, Riddell MA, Moss WJ, et al. 2006, Genotyping of measles virus in clinical specimens on the basis of oligonucleotide micro array hybridization patterns. J Clin Microbiol 44 (10) :3752-3759   DOI   ScienceOn
9 Altman DG. 1999, Practical statistics for medical research. Chapman & Hall/CRC Press, Boca Raton/London/New York/Washington, DC
10 Chizhikov V, Wagner M, Ivshina A, et al. 2002, Detection and genotyping of human group A rotaviruses by oligonucleotide microarray hybridization. J Clin Microbiol 40 (7) : 2398-2407   DOI
11 Kwok S, Higuchi R. 1989, Avoiding false positives with PCR. Nature 339: 237-238   DOI   ScienceOn
12 Saif LJ, Bohl EH, Theil KW, et al. 1980. Rotavirus-like, calicivirus-like, and 23-nm virus-like particles associated with diarrhea in young pigs. J Clin Microbiol 12 (1) : 105-111