Watanabe Mizuki, Kitamura Tomoya, Nagata Koji, Ikezawa Mitsutaka, Kameyama Ken-Ichiro, Masujin Kentaro, Kokuho Takehiro
Nippon Institute for Biological Science, Tokyo 198-0024, Japan.
Division of Transboundary Animal Disease Research, National Institute of Animal Health (NIAH), National Agriculture and Food Research Organization (NARO), Tokyo 187-0022, Japan.
Pathogens. 2023 May 29;12(6):774. doi: 10.3390/pathogens12060774.
African swine fever is a hemorrhagic viral disease with a mortality rate of nearly 100% in pigs. Hence, it is classified as a notifiable disease by the World Organization for Animal Health. Because no field-available vaccine exists, African swine fever virus (ASFV) control and eradication solely depend on good farm biosecurity management and rapid and accurate diagnosis. In this study, we developed a new indirect serological enzyme-linked immunosorbent assay (ELISA) using recombinant p11.5 protein from ASFV as a solid-phase target antigen. The cutoffs were determined by receiver operating curve analysis performed with serum samples obtained from naïve and infected pigs. Based on the results of a commercially available serological ELISA, the relative sensitivity and specificity of our assay were 93.4% and 94.4% (N = 166; area under the curve = 0.991; 95% confidence interval = 0.982-0.999), respectively. Furthermore, to compare the performance of the serological ELISAs, we conducted the assays on a panel of sera collected from pigs and boars experimentally infected with different ASFV isolates. The results indicated the greater sensitivity of the newly developed assay and its ability to detect anti-ASFV antibodies earlier after virus inoculation.
非洲猪瘟是一种出血性病毒性疾病，猪的死亡率接近100%。因此，它被世界动物卫生组织列为须通报的疾病。由于目前尚无可用于田间的疫苗，非洲猪瘟病毒（ASFV）的防控和根除完全依赖于良好的养殖场生物安全管理以及快速准确的诊断。在本研究中，我们开发了一种新的间接血清学酶联免疫吸附测定（ELISA），使用来自ASFV的重组p11.5蛋白作为固相靶抗原。通过对从未接触过病毒和感染病毒的猪的血清样本进行受试者工作特征曲线分析来确定临界值。基于市售血清学ELISA的结果，我们的检测方法的相对敏感性和特异性分别为93.4%和94.4%（N = 166；曲线下面积 = 0.991；95%置信区间 = 0.982 - 0.999）。此外，为了比较血清学ELISA的性能，我们对一组从经不同ASFV分离株实验感染的猪和公猪采集的血清进行了检测。结果表明新开发的检测方法具有更高的敏感性，并且能够在病毒接种后更早地检测到抗ASFV抗体。
