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Development of an indirect ELISA based on soluble antigen produced from virus-infected cells for detection of porcine hemagglutinating encephalomyelitis virus
Journal of Virological Methods ( IF 2.2 ) Pub Date : 2020-12-05 , DOI: 10.1016/j.jviromet.2020.114016
Chau Thi Huyen Trang , Makoto Nakanishi , Hideki Hayashidani , Takahide Taniguchi

Porcine hemagglutinating encephalomyelitis virus (PHEV) is a member of the genus Betacoronavirus and is the etiologic agent of encephalomyelitis or vomiting and wasting disease in neonatal pigs. Although there are only a few epidemiological studies that document the seroprevalence of PHEV infection, there are reports of sporadic outbreaks, including recent documentation of an influenza-like respiratory disease associated with PHEV in the United States. To address this issue, we have developed a new indirect enzyme linked immunosorbent assay (ELISA) for use in sero-epidemiological research of PHEV infection. One hundred and fifty porcine serum samples that were determined as antibody-positive or antibody-negative in virus neutralization (VN) tests were used in conjunction with PHEV-specific antigen extracted from virus-infected FS-L3 cells using RBS buffer containing 0.2 % NP-40 to develop this assay. The ELISA showed a high sensitivity (95.35 %) and specificity (96.88 %) by receiver operating characteristic (ROC) analysis, with an area under the curve (AUC) of 0.996 attesting to its accuracy. Our results revealed a strong correlation between the results of the indirect ELISA and VN test (R = 0.850, P < 0.05), with near-perfect agreement (kappa value = 0.932). These results indicate that this new indirect ELISA might be useful for diagnosis and sero-epidemiological tracking of PHEV infection.



中文翻译:

基于病毒感染细胞产生的可溶性抗原的间接ELISA的开发,用于检测猪血凝性脑脊髓炎病毒

猪血凝性脑脊髓炎病毒(PHEV)是Betacoronavirus属的成员并且是新生猪脑脊髓炎或呕吐和消瘦疾病的病原体。尽管只有很少的流行病学研究记录了PHEV感染的血清流行,但是有零星爆发的报道,包括最近在美国有关与PHEV相关的流感样呼吸道疾病的报道。为解决此问题,我们开发了一种新的间接酶联免疫吸附测定(ELISA),可用于PHEV感染的血清流行病学研究。将150份在病毒中和(VN)测试中被确定为抗体阳性或抗体阴性的猪血清样品与使用含有0.2%NP的RBS缓冲液从病毒感染的FS-L3细胞中提取的PHEV特异性抗原结合使用-40开发此测定法。ELISA显示高灵敏度(95。接收器工作特性(ROC)分析得出35%的特异性和96.88%的特异性,曲线下面积(AUC)为0.996证明了其准确性。我们的结果显示间接ELISA结果与VN测试之间存在很强的相关性(R = 0.850,P <0.05),具有接近完美的一致性(kappa值= 0.932)。这些结果表明,这种新的间接ELISA可能对PHEV感染的诊断和血清流行病学追踪有用。

更新日期:2020-12-17
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