草鱼出血病病毒抗原亚单位的研制

通过比较溶液的离子浓度对病毒形态的结构的影响，发现ＧＣＨＶ在低盐溶液盐低温（－２０℃）保存条件下，外壳蛋白会自动降解，但不十分完全，采用含１％ＮＰ４０（Ｎｏｎｉｄｅｔ－ｐ４０）的稀盐溶液处理感染病毒的细胞，经冻融及差异离心，电镜下观察到纯度较高的病毒双层衣壳，核心衣壳及散在的子粒。纯化的病毒亚单位用核酸酶消化后，与等量的福氏佐剂混合，免疫家兔制备抗体，该抗血清经台和试验测定，显示了较强的中和病毒的

Preparation of Grass Carp Hemorrhage Virus Antigen Subunit

By studying the flue structure of GCHV (Grass Carp Hemorrhage Virus), It was found that the capsid of purified GCHV can be degraded spontaneously into its morphological units by prologing in a buffer of low ionic strength, but not very completely. The virus preparation was obtained from GCHV infected cell, then treated with 1% NP40 reagent in low-salt solution and fieezed thaw ng, finally purified by differential centrifugation. Using negative staining method, out, core capsid and capsomeres of the virus were showed by electron micrograph. Afterwards, the preparation digested by RNase was equally mixed with Freund's adjuvant and immunized rabbits. The antibody was tested by ELISA and neutralization test, which demostrated the preparation can elicit good levels of antibody. The RNA-free preparation was detected by PAGE and DIG-labelled GCHV-cDNA hybridization. The sensitivity of the test is approximately to 10pg, the samples of subunit all showed negative. The preparation has a high purity, small volume and better safety. Besides, it is easy to use, also convenient in transportation and conservation. It will be a ideal vaccine against grass carp hemorrhage disease.