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Journal of Veterinary Diagnostic Investigation Vol. 20 Issue 3, 283-288
Copyright © 2008 by the American Association of Veterinary Laboratory Diagnosticians
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Full Scientific Reports

Evaluation of a nested polymerase chain reaction assay to differentiate between two genotypes of Porcine circovirus-2

Kwang Soo Lyoo, Hyeun Bum Kim and Han Soo Joo1

Correspondence: 1Corresponding Author: Han Soo Joo, University of Minnesota, 385 Animal Sci/Vet Medicine Building, 1988 Fitch Avenue, St. Paul, MN 55108, e-mail: jooxx001{at}umn.edu

Porcine circovirus-2 (PCV-2) is associated with several diseases in pigs, including postweaning multisystemic wasting syndrome (PMWS). A new genotype of PCV-2 was isolated from swine farms with and without clinical PMWS in North America. The new genotype was differentiated in a separate cluster by phylogenetic analyses and is now named PCV-2b compared with PCV-2a for the previously known genotype. The purpose of this study was to develop and evaluate a nested polymerase chain reaction (nPCR) assay to detect and differentiate between PCV-2a and PCV-2b. Genotype-specific primer sets were designed by using sequence data published for different PCV-2 strains. Specificity and sensitivity of the nPCR were examined by using PCV-2 isolates with known genotype. Nested PCR was found to be highly specific and sensitive for detecting and differentiating between the PCV-2 genotypes compared with the conventional 1-step PCR assay. Nested PCR was applied to detect PCV-2 and to identify the genotype in serum samples from swine farms with and without a clinical history of PMWS. Of 60 serum samples collected from 4 farms during clinical PMWS outbreaks, PCV-2a and PCV-2b were detected in 6 and 49 samples, respectively. Six of the 10 samples from one of the 4 farms had both PCV-2a and PCV-2b. Of 20 serum samples from 2 farms without PMWS, 11 were positive for PCV-2a only. These results suggest that the differential nPCR can be used to detect PCV-2 and to differentiate the 2 genotypes from field samples.

Key Words: Genotype differentiation • nested polymerase chain reaction • Porcine circovirus-2






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