title: A TaqMan-based real-time polymerase chain reaction for the detection of porcine parvovirus.
abstract: A real-time polymerase chain reaction (PCR) using a TaqMan probe was developed to detect porcine parvovirus (PPV). Real-time PCR was optimized to quantify PPV using a detection system (Rotor Gene 2000 detector) and a dual-labeled fluorogenic probe. The gene-specific labeled fluorogenic probe for the VP2 gene of PPV was used to detect PPV. Quantitation of PPV was accomplished by a standard curve plotting cycle threshold values (Ct) against each dilution of standard plasmids. When the specificity of the assay using specific PPV primers was evaluated by testing the PPV standard strain and other viruses, no cross-reactions were detected with non-PPV reference viruses. The detection limit of real-time PCR for PPV was 2.08log10 genome copy equivalent (gce). In this study, a real-time PCR assay was performed on 80 clinical samples and compared with a conventional PCR assay. In 48 of 80 samples, PPV DNA was detected by the conventional PCR assay. All samples positive for PPV DNA by the conventional PCR assay were also positive by the real-time PCR assay, and 12 of 32 samples that tested negative for PPV DNA by the conventional method tested positive by the real-time PCR assay. Using the real-time PCR assay, the number of samples in which PPV was detected increased by 15%. Therefore, it is considered to be a useful tool for the detection of PPV.
authors.affiliation: College of Animal Husbandry and Veterinary, Henan Agricultural University, Zhengzhou 450002, China.
authors.forename: Hong-Ying
authors.initials: HY
authors.lastname: Chen
authors.forename: Xiao-Kang
authors.initials: XK
authors.lastname: Li
authors.forename: Bao-An
authors.initials: BA
authors.lastname: Cui
authors.forename: Zhan-Yong
authors.initials: ZY
authors.lastname: Wei
authors.forename: Xin-Sheng
authors.initials: XS
authors.lastname: Li
authors.forename: Yan-Bin
authors.initials: YB
authors.lastname: Wang
authors.forename: Li
authors.initials: L
authors.lastname: Zhao
authors.forename: Zhen-Ya
authors.initials: ZY
authors.lastname: Wang
mesh_terms: D000818:Animals; D015342:DNA Probes; D004279:DNA, Viral / Q000032:analysis; D005333:Fetus / Q000821:virology; D010322:Parvoviridae Infections / Q000175:diagnosis; D028323:Parvovirus, Porcine / Q000302:isolation & purification*; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013552:Swine / Q000821:virology*; D013553:Swine Diseases / Q000175:diagnosis
chemical_list: D015342:DNA Probes; D004279:DNA, Viral