The Detection Of The Growth And Decline Rule Of Antibodies Of GPMV-NP By ELISA And The Initial Study Of The Correlation Of ELISA And HI

Goose Paramyxovirus disease named Newcastle Disease viruses of goose origin again, whichis an acute contagious disease caused by Goose Paramyxovirus(GPMV), a member of Rubulavirusfamily. Which was characerized by the lesion of digestive tract with high mortality and occurrence,and has caused a large loss in economy. Goose Paramyxovirus disease infection has spreaded inmany province in China, but attention has not been directed towards Goose Paramyxovirusinfection, so far neither Goose Paramyxovirus disease vaccine applied nor adoption of measures tocontrol of Goose Paramyxovirus disease. It has shown that Goose Paramyxovirus disease infectionhas expand in China, so it is of great interest to study the pathogen from the molecular biology. Atpresent,there are many reports about comparison of ELBA and HI detection methods of NDV ondetection technology of serology. But no report on GPMV. This test first time explores the growthand decline rule of the antibody for neucleocapsid protein by ELBA whose coating antigen isneucleocapsid protein(NP) expressed by E.coli.Neucleocapsid protein (NP) is the main structural protein of GPMV, so it has importantdiagnostic significance. So gaining NP gene and its expressing product is key to the preparation ofthe specific diagnosis antigen. The NP gene of JS/1/97/GO strain was cloned into Prokarytoicexpression vector pET-30a to construct the positive recombinant plasmid pET-30a-NP sucessfully toexpress NP. The result of SDS-PAGE analysis showed that the NP was efficiently expressed and theimmunity reactivity of the recombinant protein was confirmed by Western-blot. The purified NPprotein could be used as antigen in the development of an indirect ELISA for the detection ofantibody against NP protein of GPMV, and the optimal reactive condition were determined.Healthy gooses were vaccinated with Oil-adjvant Inactive Vaccine of goose paramyxovirus. Animalswere randomly divided into four groups. A:non-vaccinated contronl; B:the vaccinated group C:theinfected group D:the post-vaccination infected group.Harvesting the blood of gooses in 1,2,3,4,5,6,7,8,9,10,11,12,14,18 weeks respectively after vaccination or infected. We detectgrowth and decline condition of the antibody for neucleocapsid protein by ELISA whose coatingantigen is neucleocapsid protein(NP) expressed by E.coli. Comparing the linear relationship betweenthe ELISA value and the HI value of each group,Construct the regression analysis of each group. (1)the equation of the vaccinated group was ELISA=0.1981 HI+0.0541(r=0.9134, P＜0.05); (2) theequation of the infected post-vaccination group was ELISA= 0.1807 HI-0.0744 (r=0.8240, P＜ 0.05); (3) the equation of the infected group was ELISA=0.2728 HI-0.4498 (r=0.8795, P＜0.05),the result demonstrates that antibody titer detected by two methods are correlated significantly.This study only explored the relationship of two antibodies in laboratory, however, in practicalclinical detection, a great quantity of animal experiments still need to do to check if the ELISAdetection method with NP as coating antigen can replace the HI test.