title: Development of a novel TaqMan-based real-time PCR assay for the detection of porcine boca-like virus (Pbo-likeV).
abstract: The recently discovered porcine boca-like virus (Pbo-likeV) is a member of the Parvoviridae family, genus Bocavirus, and it is potentially associated with swine disease. Several studies have associated Pbo-likeV with postweaning multisystemic wasting syndrome in pigs, but the full spectrum of clinical disease and the epidemiology of Pbo-likeV infection remain unclear. The availability of rapid and reliable molecular diagnostics would aid future studies of this novel virus. Thus, we developed a sensitive and specific TaqMan-based real-time PCR assay to target the Pbo-likeV NP1 gene. The assay reproducibly detected 20 copies of a recombinant DNA plasmid containing the NP1 gene, with a dynamic range of six orders of magnitude (10(2)-10(7) copies). The assay did not cross-react with other animal viruses. Clinical evaluation found that Pbo-likeV was present in Chinese swine herds at a frequency of 44.2% (114/258). Higher infection rates were found in diseased pigs (56.1%, 101/180) compared with healthy pigs (16.7%, 13/78) (P < 0.05). Our assay for the diagnosis and quantification of Pbo-likeV was highly sensitive and specific, and should provide a reliable real-time tool for epidemiological and pathogenetic study of Pbo-likeV infection.
authors.affiliation: Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Key Laboratory of Animal Diseases, Diagnostics, and Immunology, Ministry of Agriculture, National Center for Engineering Research of Veterinary Bio-products, Nanjing 210014, Jiangsu Province, PR China.
authors.forename: Bin
authors.initials: B
authors.lastname: Li
authors.forename: Shaobo
authors.initials: S
authors.lastname: Xiao
authors.forename: Junjie
authors.initials: J
authors.lastname: Ma
authors.forename: Yanling
authors.initials: Y
authors.lastname: Liu
authors.forename: Li
authors.initials: L
authors.lastname: Mao
authors.forename: Libin
authors.initials: L
authors.lastname: Wen
authors.forename: Aihua
authors.initials: A
authors.lastname: Mao
authors.forename: Xuehan
authors.initials: X
authors.lastname: Zhang
authors.forename: Yanxiu
authors.initials: Y
authors.lastname: Ni
authors.forename: Rongli
authors.initials: R
authors.lastname: Guo
authors.forename: Junming
authors.initials: J
authors.lastname: Zhou
authors.forename: Zhengyu
authors.initials: Z
authors.lastname: Yu
authors.forename: Lixin
authors.initials: L
authors.lastname: Lv
authors.forename: Xiaomin
authors.initials: X
authors.lastname: Wang
authors.forename: Liurong
authors.initials: L
authors.lastname: Fang
authors.forename: Huanchun
authors.initials: H
authors.lastname: Chen
authors.forename: Kongwang
authors.initials: K
authors.lastname: He
mesh_terms: D000818:Animals; D052660:Bocavirus / Q000302:isolation & purification*; D019411:Clinical Laboratory Techniques / Q000379:methods; D017931:DNA Primers / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics*; D015345:Oligonucleotide Probes / Q000737:chemistry / Q000235:genetics; D010322:Parvoviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis* / Q000821:virology*; D017361:Viral Nonstructural Proteins / Q000235:genetics; D014773:Virology / Q000379:methods*
chemical_list: D017931:DNA Primers; D004279:DNA, Viral; D015345:Oligonucleotide Probes; D017361:Viral Nonstructural Proteins