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Reactivity of Porcine Epidemic Diarrhea Virus Structural Proteins to Antibodies against Porcine Enteric Coronaviruses: Diagnostic Implications

机译:猪流行性腹泻病毒结构蛋白对猪肠道冠状病毒抗体的反应性:诊断意义

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The development of porcine epidemic diarrhea virus (PEDV) antibody-based assays is important for detecting infected animals, confirming previous virus exposure, and monitoring sow herd immunity. However, the potential cross-reactivity among porcine coronaviruses is a major concern for the development of pathogen-specific assays. In this study, we used serum samples (n = 792) from pigs of precisely known infection status and a multiplex fluorescent microbead-based immunoassay and/or enzyme-linked immunoassay platform to characterize the antibody response to PEDV whole-virus (WV) particles and recombinant polypeptides derived from the four PEDV structural proteins, i.e., spike (S), nucleocapsid (N), membrane (M), and envelope (E). Antibody assay cutoff values were selected to provide 100% diagnostic specificity for each target. The earliest IgG antibody response, mainly directed against S1 polypeptides, was observed at days 7 to 10 postinfection. With the exception of nonreactive protein E, we observed similar antibody ontogenies and patterns of seroconversion for S1, N, M, and WV antigens. Recombinant S1 provided the best diagnostic sensitivity, regardless of the PEDV strain, with no cross-reactivity detected against transmissible gastroenteritis virus (TGEV), porcine respiratory coronavirus (PRCV), or porcine deltacoronavirus (PDCoV) pig antisera. The WV particles showed some cross-reactivity to TGEV Miller and TGEV Purdue antisera, while N protein presented some cross-reactivity to TGEV Miller. The M protein was highly cross-reactive to TGEV and PRCV antisera. Differences in the antibody responses to specific PEDV structural proteins have important implications in the development and performance of antibody assays for the diagnosis of PEDV enteric disease.
机译:基于猪流行性腹泻病毒(PEDV)抗体的检测方法的开发对于检测受感染的动物,确认先前的病毒暴露以及监测母猪的免疫力非常重要。然而,猪冠状病毒之间潜在的交叉反应性是病原体特异性检测方法发展的主要关注点。在这项研究中,我们使用了精确知道感染状况的猪的血清样品( n = 792)和基于多重荧光微珠的免疫测定和/或酶联免疫测定平台来表征对PEDV的抗体反应源自四个PEDV结构蛋白的全病毒(WV)颗粒和重组多肽,即刺突(S),核衣壳(N),膜(M)和包膜(E)。选择抗体测定的临界值以为每个靶标提供100%的诊断特异性。在感染后第7至10天观察到了主要针对S1多肽的最早IgG抗体反应。除非反应性蛋白E外,我们观察到了相似的抗体基因组以及S1,N,M和WV抗原的血清转化模式。不论PEDV株如何,重组S1均可提供最佳的诊断敏感性,且未检测到与可传播的胃肠炎病毒(TGEV),猪呼吸道冠状病毒(PRCV)或猪三角洲冠状病毒(PDCoV)猪抗血清的交叉反应。 WV颗粒对TGEV Miller和TGEV Purdue抗血清具有一定的交叉反应性,而N蛋白对TGEV Miller具有一定的交叉反应性。 M蛋白与TGEV和PRCV抗血清高度交叉反应。抗体对特定PEDV结构蛋白反应的差异对诊断PEDV肠道疾病的抗体检测方法的开发和性能具有重要意义。

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