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.
Subject(s)
Animal Husbandry , Chromatography, Affinity/methods , Encephalitis Virus, Japanese/immunology , Serologic Tests/methods , Surveys and Questionnaires , Sus scrofa/immunology , Sus scrofa/virology , Animals , Blotting, Western , Cloning, Molecular , DNA, Complementary/genetics , Encephalitis Virus, Japanese/isolation & purification , Geography , Protein Structure, Tertiary , Protein Subunits/isolation & purification , Reagent Kits, Diagnostic , Recombinant Proteins/metabolism , Republic of Korea , Viral Proteins/chemistry , Viral Proteins/metabolismABSTRACT
OBJECTIVES: Cases of human brucellosis in Korea have recently increased due to the increasing incidence of bovine brucellosis. The authors conducted this study to elucidate the status of brucellosis through seroepidemiologic study. METHODS: We selected our study population from a high risk group. We conducted a questionnaire survey and obtained blood samples to determine the seroprevalence of brucellosis antibodies for 10 days in February, 2005. The titers of brucellosis were measured by the combination of standard tube agglutination test (STA) and enzyme-linked immunosorbent assay (ELISA) test. RESULTS: Our study subjects comprised 1,075 cases: 971 livestock workers, 51 veterinarians, and 53 artificial inseminators. In the STA test, 27 cases (2.5%) had titers of greater than or equal to 1:20. Of 1,068 cases (7 cases were excluded due to previous brucellosis), 7 cases of brucellosis were diagnosed with titers of 1:160, giving a seroprevalence of brucellosis of 0.66%. The seroprevalence in the male group was 0.95%, and that of livestock workers, veterinarians, and artificial inseminators was 0.52%, 4.17%, and 0.00%, respectively. The Spearman's correlation coefficient between the positive rate of bovine brucellosis per capita and household and human brucellosis was 0.806 and 0.744, respectively. The concordance rate between the Korea National Institute of Health and the Gyeongsangbuk-do Institute of Health and Environment by the STA and ELISA tests was 94.7% and 100.0%, respectively. CONCLUSIONS: The study results indicated in higher seroprevalence rate among veterinarians than among livestock workers and artificial inseminators. Because veterinarians may be exposed to this high risk, effective working guidelines for veterinarians to guard against brucellosis must be developed. Moreover, more extensive epidemiologic research for laboratory workers and meat handlers is needed.
