行政院國家科學委員會專題研究計畫成果報告:應用S1表現蛋白偵測傳染性支氣管炎病毒抗體

Enzyme-linked immnunosorbent assay
(ELISA) using an expression S protein
fragment as an antigen was developed for
detection of antibodies against IBV. The
primers (IBVc1-IBVc2) were designed
according to the published entire S1
nucleotide sequence of Taiwan IBV isolate
Tw/1171/92. The F frgament of S1 gene was
amplified using primers IBVc1-IBVc2,
cloned into the pRSET vectors and
transformed into competent E. coli BL21
(DE3). A 27.5 kD S1-F fusion protein was
successfully expressed, affinity-purified and
used as a coated antigen for serum antibody
detection by ELISA. Although there was
little cross-reactivity in the serum
neutralization (SN) test, cross-reactivity was
observed in this ELISA system among
different IBV strains, H120, Taiwan Group I
and Taiwan Group II. The ELISA technique
using expression S1-F protein is able to
detect antibodies from different IBV
serotypes and appears to be suitable for
large-scale serological surveys.