Abstract
Among vertebrate species, pigs are a major amplifying host of Japanese encephalitis virus (JEV) and measuring their seroconversion is a reliable indicator of virus activity. Traditionally, the hemagglutination inhibition test has been used for serological testing in pigs; however, it has several limitations and, thus, a more efficient and reliable replacement test is required. In this study, we developed a new immunochromatographic test for detecting antibodies to JEV in pig serum within 15 min. Specifically, the domain III region of the JEV envelope protein was successfully expressed in soluble form and used for developing the immunochromatographic test. The test was then applied to the surveillance of Japanese encephalitis (JE) in Korea. We found that our immunochromatographic test had good sensitivity (84.8%) and specificity (97.7%) when compared with an immunofluorescence assay used as a reference test. During the surveillance of JE in Korea in 2012, the new immunochromatographic test was used to test the sera of 1,926 slaughtered pigs from eight provinces, and 228 pigs (11.8%) were found to be JEV-positive. Based on these results, we also produced an activity map of JEV, which marked the locations of pig farms in Korea that tested positive for the virus. Thus, the immunochromatographic test reported here provides a convenient and effective tool for real-time monitoring of JEV activity in pigs.
Highlights
Japanese encephalitis virus (JEV), belonging to the genus Flavivirus in the family Flaviviridae, is a causative pathogen of acute encephalitis in humans [1]
In order to improve the surveillance of JE, we developed an immunochromatographic test for detecting antibodies to JEV in pigs and successfully applied it in national JE surveillance conducted in South Korea
Gel electrophoresis and western blotting analyses showed that the over-expressed protein was highly purified in native conditions and reacted strongly with JEV-positive human sera (Fig 3)
Summary
Japanese encephalitis virus (JEV), belonging to the genus Flavivirus in the family Flaviviridae, is a causative pathogen of acute encephalitis in humans [1]. An Immunochromatograhic Test for Detecting Antibodies to JEV in Pig regions at the 50 and 30 ends [1]. JEV is transmitted via a cycle that includes (a) infection of pigs and water birds by horizontal transmission from mosquito vectors, (b) infection of certain mosquitoes by horizontal transmission from vertebrate amplifiers, and (c) vertical transmission from adult female mosquitoes to their progeny [2]. Humans and horses are infected incidentally in this cycle and considered dead-end hosts because the level of viremia is insufficient to infect the mosquito vector. According to the latest report in 2011 [3], more than 67,000 cases of Japanese encephalitis (JE), with 20–30% fatality, occur annually throughout Asia and the Pacific regions
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