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利用蔗糖密度梯度离心技术从自然发病的栉孔扇贝 (Chlamysfarrreri)组织分离纯化急性病毒性坏死症(Acutevirusnecrobioticdisease,AVND)病毒 ,并以此为抗原免疫Balb c小鼠。将免疫小鼠的脾细胞与NS_1骨髓瘤细胞进行融合 ,最终筛选出 4株能稳定传代并分泌该病毒特异性单克隆抗体 (MonoclonalAntibody ,MAb)的杂交瘤细胞系。应用胶体金标记免疫电镜技术在超微水平上对这 4株MAbs进行测定表明 ,它们对AVND病毒均具有高度的特异性 ,并且所识别的特异性位点均位于病毒粒子囊膜的纤突上。应用该单克隆抗体对不同养殖季节的一龄贝进行间接ELISA检测发现 ,7月中旬至 7月底病毒感染率与感染强度均处于当年的最高峰 ,与这一时期栉孔扇贝所表现出的最高死亡率完全吻合
Acutevirus necrobiotic disease (AVND) virus was isolated and purified from the naturally infected Chlamys farreri tissue by sucrose density gradient centrifugation and used as antigen to immunize Balb c mice. The spleen cells of immunized mice were fused with NS1 myeloma cells, and finally 4 hybridoma cell lines which could stably passage and secrete this virus specific monoclonal antibody (Monoclonal Antibody, MAb) were screened out. The application of colloidal gold-labeled immunoelectron microscopy to the determination of these 4 MAbs at the ultrasmall level showed that they all have high specificity for the AVND virus and the identified specific sites are located on the fibroblasts of the viral particle envelope . Using this monoclonal antibody for indirect detection of first instar in different seasons, it was found that the virus infection rate and the intensity of infection were all the highest in the same year from mid July to the end of July, and the highest in this period Mortality is exactly the same