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首页> 外文期刊>Journal of Biotechnology >Expression of the porcine circovirus type 2 capsid protein subunits and application to an indirect ELISA
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Expression of the porcine circovirus type 2 capsid protein subunits and application to an indirect ELISA

机译：猪圆环病毒2型衣壳蛋白亚基的表达及其在间接ELISA中的应用

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Porcine circovirus type 2 (PCV2) is considered to be associated with post-weaning multisystemic wasting syndrome (PMWS), which is a newly emerged economically important swine disease. The entire coding region of open reading frame 2 (ORF2), encoding the viral capsid protein (Cap), of PCV2 was cloned and sequenced from the clinical specimen obtained from PMWS-affected piglets. Six recombinant subunits, A-F, spanning the defined regions of Cap were produced by E. coli expression system and used as antigens for testing their reactivities with swine sera in the indirect ELISA. The recombinant Cap subunit-based ELISA was evaluated by examining a panel of 12 PCV2-negative and 26 PCV2-positive sera. When the positiveegative cut-off value was set at the mean value of negative sera plus 3 standard deviations, all subunits-based ELISA demonstrated 100% specificities. The N-terminal subunits, A and B, revealed poor reactivity with positive swine sera, whereas, greater immunoreactivity was observed for the C-terminal subunits of which subunits C and D demonstrated good sensitivities of 96.2% and 84.6%, respectively. The recombinant Cap subunits possessing defined antigenicity are easy to produce and the subunit-based ELISA was developed with a high specificity and sensitivity that may provide a useful method for routine serodiagnosis of PCV2 infection.
机译：猪圆环病毒2型（PCV2）被认为与断奶后多系统消耗综合症（PMWS）有关，这是一种新兴的在经济上重要的猪病。从得自受PMWS影响的仔猪的临床标本中克隆并测序PCV2的开放阅读框2（ORF2）的整个编码区，并编码病毒衣壳蛋白（Cap）。大肠杆菌表达系统产生了跨越Cap限定区域的六个重组亚基A-F，并用作抗原，用于在间接ELISA中测试其与猪血清的反应性。通过检查一组12个PCV2阴性和26个PCV2阳性血清来评估基于重组Cap亚基的ELISA。当将阳性/阴性截止值设置为阴性血清的平均值加3个标准差时，所有基于亚基的ELISA都显示出100％的特异性。 N末端亚基A和B显示出与阳性猪血清的不良反应性，而C末端亚基C和D分别显示出良好的敏感性，分别为96.2％和84.6％。具有确定的抗原性的重组Cap亚基易于生产，并且以高特异性和灵敏度开发了基于亚基的ELISA，这可能为PCV2感染的常规血清诊断提供了有用的方法。

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