title: A new nanoPCR molecular assay for detection of porcine bocavirus.
abstract: Nanoparticle-assisted polymerase chain reaction (nanoPCR) is a novel method for the rapid amplification of DNA and has been used for the detection of virus. For detection of porcine bocavirus (PBoV), a sensitive and specific nanoPCR assay was developed with a pair of primers that were designed based on NS1 gene sequences available in GenBank. Under the optimized conditions of the PBoV nanoPCR assay, the nanoPCR assay was 100-fold more sensitive than a conventional PCR assay. The lower detection limit of the nanoPCR assay was about 6.70×10(1) copies. The nanoPCR assay amplified the specific 482-bp fragment of the PBoV NS1 recombinant plasmid but did not produce any product with genomic DNA or cDNA of porcine parvovirus, porcine circovirus type II, porcine reproductive and respiratory syndrome virus, pseudorabies virus, classic swine fever virus, Encephalomyocarditis virus, Porcine Teschovirus or African swine fever virus plasmid. Of 65 clinical samples collected from diseased pigs, 73.8% and 86.2% were determined to be PBoV positive by PBoV conventional PCR and PBoV nanoPCR assay, respectively. Of 36 clinical samples from healthy pigs, 27.8% and 44.4% were PBoV positive by PBoV conventional PCR and PBoV nanoPCR assay, respectively. The nanoPCR assay will be useful for diagnosing PBoV and for studying its epidemiology and pathology.
authors.affiliation: Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, China.
authors.forename: Xiaoling
authors.initials: X
authors.lastname: Wang
authors.affiliation: Foshan University, Foshan, Guandong, China. Electronic address: baiaiquan2008@163.com.
authors.forename: Aiquan
authors.initials: A
authors.lastname: Bai
authors.affiliation: Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, China.
authors.forename: Miaomiao
authors.initials: M
authors.lastname: Kong
authors.affiliation: Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, China.
authors.forename: Yuchao
authors.initials: Y
authors.lastname: Cui
authors.affiliation: Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, China.
authors.forename: Xingjie
authors.initials: X
authors.lastname: Ma
authors.affiliation: College of Animal Science and Veterinary Medicine, Tianjin Agricultural University, Tianjin, China.
authors.forename: Xia
authors.initials: X
authors.lastname: Ai
authors.affiliation: Nanyang Normal University, 1638 Wolong Road, Nanyang 473061, China. Electronic address: qinghaitang109@126.com.
authors.forename: Qinghai
authors.initials: Q
authors.lastname: Tang
authors.affiliation: Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, China. Electronic address: cuishangjin@126.com.
authors.forename: Shangjin
authors.initials: S
authors.lastname: Cui
mesh_terms: D000818:Animals; D052660:Bocavirus / Q000235:genetics* / Q000302:isolation & purification*; D017931:DNA Primers / Q000235:genetics; D010322:Parvoviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis* / Q000821:virology*; D017361:Viral Nonstructural Proteins / Q000235:genetics
chemical_list: D017931:DNA Primers; D017361:Viral Nonstructural Proteins
keywords: Detection; Epidemic characterization; NS1 gene; NanoPCR; PBoV