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基于截断可溶性 ORF2 蛋白的 ELISA 检测试剂盒用于检测猪圆环病毒 2 型抗体。

An ELISA based on a truncated soluble ORF2 protein for the detection of PCV2 antibodies in domestic pigs.

机构信息

State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Animal Virology of Ministry of Agriculture, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China.

出版信息

Virol Sin. 2010 Jun;25(3):191-8. doi: 10.1007/s12250-010-3085-5. Epub 2010 Jun 6.

Abstract

Postweaning multisystemic wasting syndrome (PMWS) is an important swine disease that is closely associated with porcine circovirus type 2 (PCV2). The capsid protein (Cap protein) is a major structural protein that has at least three immunoreactive regions, and it can be a suitable candidate antigen for detecting the specific antibodies of a PCV2 infection. In the present study, an indirect enzyme-linked immunosorbent assay (TcELISA) based on a truncated soluble Cap protein produced in Escherichia coli (E.coli) was established and validated for the diagnostic PCV2 antibodies in swine. The TcELISA was validated by comparison with an indirect immunofluorescence assay (IIFA). The diagnostic sensitivity (DSN), specificity (DSP), and accuracy of the TcELISA were 88.6%, 90.7% and 89.4%, respectively. The agreement rate was 89.38% between results obtained with TcELISA and IIFA on 113 field sera. A cross-reactivity assay showed that the method was PCV2-specific by comparison with other sera of viral disease. Therefore,the TcELISA will be helpful for the development of a reliable serology diagnostic test for large scale detection of PCV2 antibodies and for the evaluation of vaccine against PCV2 in swine.

摘要

断奶后多系统消耗综合征(PMWS)是一种重要的猪病,与猪圆环病毒 2 型(PCV2)密切相关。衣壳蛋白(Cap 蛋白)是一种主要的结构蛋白,至少有三个免疫反应区,它可以作为检测 PCV2 感染特异性抗体的合适候选抗原。本研究建立并验证了基于大肠杆菌(E.coli)中表达的截短可溶性 Cap 蛋白的间接酶联免疫吸附试验(TcELISA),用于检测猪 PCV2 抗体。该 TcELISA 通过与间接免疫荧光试验(IIFA)进行比较来验证。该 TcELISA 的诊断灵敏度(DSN)、特异性(DSP)和准确性分别为 88.6%、90.7%和 89.4%。在 113 份现场血清中,用 TcELISA 和 IIFA 检测结果的符合率为 89.38%。交叉反应试验表明,与其他病毒性疾病的血清相比,该方法具有 PCV2 特异性。因此,该 TcELISA 将有助于开发一种可靠的血清学诊断检测方法,用于大规模检测 PCV2 抗体,并评估针对 PCV2 的疫苗在猪中的效果。

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