胶体金免疫层析法检测猪PRVgE抗体方法的建立及应用

Establishment and application of colloidal gold immuno-chromatographicassay for detecting gE antibody of swine pseudo rabies virus

利用PCR技术从猪伪狂犬病毒基因组中克隆gE抗原表位基因,并将其插入到载体pET-22b(+)中,构建成原核表达质粒pET-22b(+)-gE,使其在E.coli BL21(DE3)中以IPTG诱导表达,经斑点杂交证实表达产物具有抗原性.表达产物纯化后作为抗原,结合胶体金标记技术,运用双抗原夹心法于国内首先建立了猪PRVgE抗体检测的免疫层析试纸条.该方法操作简单灵敏度高、特异性好,适合猪伪狂犬的临床诊断.

By PCR,a gE antigen epitope gene was obtained from the genome of swine pseudo rabies virus. We inserted the gene into an expression vector, ,to yield the recombinant plasmid pET-22b（＋）-gE. After induced by IPTG, a high expression of gE protein was obtained. The product of expression was specific to antisera against PRV by protein dot blot analysis. After purified, the gene-expressed gE acts as attenuated PRV antigen.Based-on sandwich ELISA technique, immunochromatographicassay test paper for detecting PRV is firstly developed in China. The method is more sensitive and specific than other methods and fit for clinical diagnosis of PRV.