Preparation of grass carp hemorrhage virus antigen subunit
Abstract: 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.