Journal
JOURNAL OF VIROLOGICAL METHODS
Volume 187, Issue 1, Pages 15-19Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jviromet.2012.06.024
Keywords
Porcine parvovirus (PPV); Porcine circovirus type 2 (PCV-2); Duplex real-time PCR; SYBR-Green
Funding
- Innovation Scientists and Technicians troop Construction Projects of Zhengzhou City, Henan, China [10XTD148]
- Key Sci-tech Project of Zhengzhou City, Henan, China [0910SGYN12319-5]
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The development of a SYBR Green-based duplex real-time PCR is described for simultaneous detection of porcine parvovirus (PPV) and porcine circovirus type 2 (PCV-2) genomes. Viral genomes were identified in the same sample by their distinctive melting temperature (T-m) which is 77.5 degrees C for PPV VP2 313 bp amplicon and 82.3 degrees C for PCV-2 ORF2 171 bp amplicon, respectively. The detection limit of the method was 0.01 TCID50/mL for PPV and PCV-2, about 10 times more sensitive than conventional PCR. In addition, PPV and PCV-2 viral load were measured in 126 field samples, confirming the sensitivity and specificity, and the result showed that 70/126 samples were positive for PPV and 92/126 samples were positive for PCV2 by the duplex real-time PCR. This method may be a useful alternative rapid and reliable method for the detection of PPV/PCV-2 co-infection. (C) 2012 Elsevier B.V. All rights reserved.
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