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Published Tuesday, January 01, 2013: Journal of Virological Methods - Volume 187, Issue 1, January 2013, Pages 15–19
Simultaneous Detection of Porcine Parvovirus And Porcine Circovirus Type 2 By Duplex Real-Time PCR And Amplicon Melting Curve Analysis using SYBR Green
Lan-lan Zheng, Ya-bin Wang, Ming-feng Li, Hong-ying Chen, Xian-po Guo, Jing-wei Geng, Zhen-ya Wang, Zhan-yong Wei, Bao-an Cui
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 (Tm) which is 77.5ºC for PPV VP2 313 bp amplicon and 82.3ºC for PCV-2 ORF2 171 bp amplicon, respectively. The detection limit of the method was 0.01TCID50/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.


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