Development of a Novel Indirect ELISA for the Serological Diagnosis of African Swine Fever Using p11.5 Protein as a Target Antigen
Mizuki Watanabe,
Tomoya Kitamura,
Koji Nagata,
Mitsutaka Ikezawa,
Ken-ichiro Kameyama,
Kentaro Masujin,
Takehiro Kokuho
Affiliations
Mizuki Watanabe
Nippon Institute for Biological Science, Tokyo 198-0024, Japan
Tomoya Kitamura
Division of Transboundary Animal Disease Research, National Institute of Animal Health (NIAH), National Agriculture and Food Research Organization (NARO), Tokyo 187-0022, Japan
Koji Nagata
Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo 113-8657, Japan
Mitsutaka Ikezawa
Division of Transboundary Animal Disease Research, National Institute of Animal Health (NIAH), National Agriculture and Food Research Organization (NARO), Tokyo 187-0022, Japan
Ken-ichiro Kameyama
Division of Transboundary Animal Disease Research, National Institute of Animal Health (NIAH), National Agriculture and Food Research Organization (NARO), Tokyo 187-0022, Japan
Kentaro Masujin
Division of Transboundary Animal Disease Research, National Institute of Animal Health (NIAH), National Agriculture and Food Research Organization (NARO), Tokyo 187-0022, Japan
Takehiro Kokuho
Division of Transboundary Animal Disease Research, National Institute of Animal Health (NIAH), National Agriculture and Food Research Organization (NARO), Tokyo 187-0022, Japan
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.
