J Vet Sci.  2006 Sep;7(3):271-275. 10.4142/jvs.2006.7.3.271.

Development and evaluation of indirect ELISA for the detection of antibodies against Japanese encephalitis virus in swine

• Yang DK
• Kim BH
• Lim SI
• Kwon JH
• Lee KW
• Choi CU
• Kweon CH

• KMID: 1089912
• DOI: http://doi.org/10.4142/jvs.2006.7.3.271

Abstract

The Japanese encephalitis virus (JEV) is one of causative agents of reproductive failure in pregnant sows. An indirect enzyme-linked immunosorbent assay (I-ELISA) was examined for its potential use in the rapid monitoring of the JEV, and the results were compared with those from the hemagglutination inhibition (HI) and serum neutralization (SN) tests. The comparative analysis showed that the results of I-ELISA showed a significant correlation with the conventional HI (r = 0.867) and SN tests (r = 0.804), respectively. When the I-ELISA results were compared with the traditional diagnostic assays, the sensitivity of the I-ELISA was 94.3% with the HI test and 93.7% with the SN test, respectively. The specificity was found to be 81.4% and 80.0% with the HI and SN tests, respectively. To determine the applicability of I-ELISA in the field, the serum samples from 720 pigs were collected from 4 regions in Korea between July and August 2004. The results indicated that 21.7% of screened pigs were seropositive for the JEV. The seropositive rates of JEV in the 4 provinces were 12.6% in Gyeonggi, 45.0% in Gyeongnam, 16.7% in Jeonbuk, and 12.2% in Jeju. The I-ELISA methodology developed in this study was shown to have considerable sensitivity and specificity through a comparison with HI and the SN tests. Therefore, it might be one of convenient methods for screening a large number of samples in various fields.

Keyword

diagnostic efficiency; I-ELISA; Japanese encephalitis virus; JEV

MeSH Terms

Animals
Antibodies, Viral/blood
Antigens, Viral/immunology
Encephalitis Virus, Japanese/immunology/*isolation&purification
Encephalitis, Japanese/blood/immunology/*veterinary/virology
Enzyme-Linked Immunosorbent Assay/methods/*veterinary
Female
Hemagglutination Inhibition Tests/veterinary
Korea
Neutralization Tests/veterinary
Swine
Swine Diseases/blood/immunology/*virology

Figure

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  Fig. 1 Titration of the JEV antigen by I-ELISA using different concentrations of the purified JEV. The numbers show the HI titers of the pig sera.

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  Fig. 2 Titration of the serum by I-ELISA. The antigen was diluted to 1 : 40 (1.25 µg/ml) in 1% skim milk. The numbers show the HI titers of the pig sera.

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  Fig. 3 Comparison of the I-ELISA and HI tests for the JEV antibodies with the positive (105) and negative (27) sow sera.

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  Fig. 4 Comparison of indirect ELISA and SN test for the JEV anribodies with 132 sow sera obtained from 6 farms.

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  Fig. 5 Seroprevalence of the JEV from 720 pigs in 2004.

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