title: Simultaneous detection of porcine parvovirus and porcine circovirus type 2 by duplex real-time PCR and amplicon melting curve analysis using SYBR Green.
abstract: 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°C for PPV VP2 313bp amplicon and 82.3°C for PCV-2 ORF2 171bp amplicon, respectively. The detection limit of the method was 0.01TCID(50)/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.
authors: College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou, Henan, China.
authors: Lan-lan
authors: LL
authors: Zheng
authors: Ya-bin
authors: YB
authors: Wang
authors: Ming-feng
authors: MF
authors: Li
authors: Hong-ying
authors: HY
authors: Chen
authors: Xian-po
authors: XP
authors: Guo
authors: Jing-wei
authors: JW
authors: Geng
authors: Zhen-ya
authors: ZY
authors: Wang
authors: Zhan-yong
authors: ZY
authors: Wei
authors: Bao-an
authors: BA
authors: Cui
mesh_terms: D000818:Animals; D052160:Benzothiazoles; D018173:Circoviridae Infections / Q000175:diagnosis / Q000662:veterinary / Q000821:virology; D017925:Circovirus / Q000235:genetics*; D060085:Coinfection; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis; D003959:Diamines; D057230:Limit of Detection; D009691:Nucleic Acid Denaturation; D009930:Organic Chemicals; D010322:Parvoviridae Infections / Q000175:diagnosis / Q000662:veterinary / Q000821:virology; D028323:Parvovirus, Porcine / Q000235:genetics*; D011804:Quinolines; D060888:Real-Time Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis / Q000821:virology; D019562:Viral Load
chemical_list: D052160:Benzothiazoles; D017931:DNA Primers; D004279:DNA, Viral; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I