Japanese Journal of Veterinary Research;Volume 32, Number 2

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LABELED AVIDIN-BIOTIN ENZYME-LINKED IMMUNOSORBENT ASSAY (LAB-ELISA) FOR DETECTION OF JAPANESE ENCEPHALITIS ANTIBODY IN SWINE SERA

CHANG, Hsi-Chi;OHKUBO, Yukihiro;TAKASHIMA, Ikuo;ARIKAWA, Jiro;HASHIMOTO, Nobuo

Permalink : http://hdl.handle.net/2115/4693
KEYWORDS : LAB-ELISA;avidin-biotin;Japanese encephalitis;swine

Abstract

Labeled avidin-biotin (LAB) enzyme-linked immunosorbent assay (ELISA) was applied to detect Japanese encephalitis (JE) antibody in swine sera. Purified JaGAr-01 vaccine of JE, biotin labeled anti-swine Ig or IgM rabbit IgG conjugate and horseradish peroxidase (HRP) were employed in the assay. The μ chain specific anti-IgM conjugate could detect only IgM antibody, but the anti-Ig conjugate could detect both IgM and IgG antibodibes to JE. When 2-2'-azino di-[3-ethylbenzthiazoline sulphonic acid](ABTS) was used as a substrate to HRP, sensitivity of LAB-ELISA was essentially the same as that of ordinary ELISA, but approximately 20 times higher than that of hemagglutination inhibition (HI) test. LAB-ELISA titers with ABTS were approximately 5 times higher than those with 5-amino salcylic acid (5-AS). However, non-specific reactions of sera were about 4 times lower in LAB-ELISA than in ordinary ELISA.

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