Observation Of Morphology On Immune Animals By Cystic Fluid Of Cysticercus Pisiformis And Research Of Rappid Diagnosis Method

Cysticercosis Pisiformis in Rabbit is a multiple and common cestodiasis that is causedby Cysticercus pisiformis parasitizing rabbits’liver、omentum majus、mesenterium and abdominalcavity. But there has been no a diagnosis method about living rabbit so far. In order to establish arapid and sensitive ELISA method, the research was to verify the immunogenicity andreactogenicity of C. pisiformis cystic fluid by the immune morphology changes of experimentalanimals and PVC-Dot-ELISA method severally. It was to validate the feasibility of C. pisiformis’cystic fluid that can be used as immunogen. The cystic fluid of C. pisiformis was purified、screened and appraised. It is to establish the blocking ELISA method with mice’s positive serumsby immunization method. The established blocking ELISA was used to detect the antibody ofEncephalitozoonosis in Rabbit、Viral Haemorrhagic Disease in Rabbit、Pasteurellosis in Rabbit、Toxoplasmosis in Rabbit and Coccidiosis in Rabbit for the specific indentification. The clinicalsamples were detected by the blocking ELISA to get the detection rate.The results of immune morphology changes showed that the number of splenic noduleincreased, the germinal center developed obviously and the periarterial lymphatic sheathconcentrated. The number of T-lymphocyte and plasmacyte in laminae propria of ileum andmesenteric lymph nodes increased. There were more positive T-lymphocyte and plasmacyte inspleen and ielum of mice. It indicated that the experimental animals had different levels ofimmune response. The PVC-Dot-ELISA method results showed that the serums of six mice allhave effective titer. The results showed that cystic fluid which is above50kDa is interest protein.The interest protein had a high density. The analysis of SDS-PAGE showed that the molecular weight of interest protein was in the vicinity of63kDa which was consistent with the expectation.The blocking ELISA method results showed that the optimum concentration of coating antigenand the optimum dilution rate of first antibody for ELISA are5μg/mL and1:800respectively. Theoptimum coating time for ELISA was staying overnight at4℃. The blocking condition for ELISAwas5%skimmed milk incubation at37℃for3h. The optimum dilution rate and working time ofsecond antibody were1:5000and incubation at37℃for30min. The specificity detection resultsshowed there was no cross reactivity among Encephalitozoonosis in Rabbit、Viral HaemorrhagicDisease in Rabbit、Pasteurellosis in Rabbit、Toxoplasmosis in Rabbit and Coccidiosis in Rabbit.One hundred clinical samples were detected by the blocking ELISA and the detection rate was82.35%. The investigation verified that C. pisiformis’ cystic fluid has good immunogenicity andreactogenicity. The experimental animal not only had humoral immune response and cellularimmunity, but also produced positive serums. The established blocking ELISA method had highspecificity and sensibility.