Detection of porcine parvovirus using a taqman-based real-time pcr with primers and probe designed for the NS1 gene.
A TaqMan-based real-time polymerase chain reaction (PCR) assay was devised for the detection of porcine parvovirus (PPV). Two primers and a TaqMan probe for the non-structural protein NS1 gene were designed. The detection limit was 1 x 10² DNA copies/μL, and the assay was linear in the range of 1 x 10² to 1 x 10⁹ copies/μL. There was no cross-reaction with porcine circovirus 2 (PCV2), porcine reproductive and respiratory syndrome virus (PRRSV), pseudorabies virus (PRV), classical swine fever virus (CSFV), or Japanese encephalitis virus (JEV). The assay was specific and reproducible. In 41 clinical samples, PPV was detected in 32 samples with the real-time PCR assay and in only 11 samples with a conventional PCR assay. The real-time assay using the TaqMan-system can therefore be practically used for studying the epidemiology and management of PPV.
Division of Swine Infectious Disease, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of CAAS, Harbin, China 150001.
Cuiping
C
Song
Chao
C
Zhu
Chaofan
C
Zhang
Shangjin
S
Cui
D000818:Animals; D017931:DNA Primers / Q000737:chemistry; D015342:DNA Probes / Q000737:chemistry; D006801:Humans; D028323:Parvovirus, Porcine / Q000235:genetics*; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis / Q000821:virology; D017361:Viral Nonstructural Proteins / Q000235:genetics*
D017931:DNA Primers; D015342:DNA Probes; C106202:NS1 protein, parvovirus; D017361:Viral Nonstructural Proteins