Animal and Plant Quarantine Agency, MAFRA, Anyang 430-757, Republic of Korea; College of Veterinary Medicine and Research Institute for Veterinary Science, Seoul National University, Seoul 151-742, Republic of Korea.
J Virol Methods. 2013 Dec;194(1-2):26-32. doi: 10.1016/j.jviromet.2013.07.030. Epub 2013 Aug 15.
Porcine circovirus type 2 (PCV2) is the causative agent of postweaning multisystemic wasting syndrome (PMWS) in swine. Although the incidences of PCV2-related diseases are ubiquitous throughout the world, the serological tools are rather limited, mainly because the virus does not induce any cytopathic effects in cells. The purpose of this study was to develop a rapid, sensitive and easy quantitative immunofluorescence assay (QIFA) using the recombinant PCV2 nucleocapsid protein (NCP) for the detection of PCV2-specific antibodies in pig sera. The recombinant PCV2 NCP was expressed in Vero cells by a lentivirus system. The performance of QIFA using these Vero cells as a diagnostic antigen was compared with currently available C-ELISA and I-ELISA; the relative sensitivity turned out to range from 92.5% up to 99.3%. The relative specificity was 93.3% when compared to C-ELISA as the gold standard. The serological experiment also indicated the inverse relationship between QIFA and the viral load in serum, semen, feces samples from 7 PCV2-positive boars. In addition, the PCV2 sequence detected from bone marrow cells shows 99% of sequence identity with PCV2 genome, confirming the infectivity of PCV2.
猪圆环病毒 2 型(PCV2)是引起猪断奶后多系统衰弱综合征(PMWS)的病原体。尽管 PCV2 相关疾病的发病率在全球范围内普遍存在,但血清学工具相当有限,主要是因为该病毒不会在细胞中引起任何细胞病变效应。本研究旨在开发一种快速、敏感和易于定量的免疫荧光检测(QIFA)方法,使用重组 PCV2 核衣壳蛋白(NCP)检测猪血清中的 PCV2 特异性抗体。重组 PCV2 NCP 通过慢病毒系统在 Vero 细胞中表达。使用这些 Vero 细胞作为诊断抗原的 QIFA 性能与当前可用的 C-ELISA 和 I-ELISA 进行了比较;相对灵敏度范围从 92.5%到 99.3%。与作为金标准的 C-ELISA 相比,相对特异性为 93.3%。血清学实验还表明,QIFA 与 7 头 PCV2 阳性公猪血清、精液、粪便样本中的病毒载量呈反比关系。此外,从骨髓细胞中检测到的 PCV2 序列与 PCV2 基因组的序列同一性为 99%,证实了 PCV2 的感染性。