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《Chinese Journal of Veterinary Science》 2008-09
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Development of the detection system for infectious bursal disease virus

WANG Zhong-can1,WANG Yong-shan2,TANG Yu-de1,TAN Wei-guo1,SHI Zheng-liang1,OUYANG wei2,3(1.Huadong Research Institute of Medical Biotechnics,Center for Disease Control and Prevention of Nanjing Command,Nanjing 210002,China;2 Institute of Veterinary Medicine,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China;3 College of Veterinary Medicine,Nanjing Agricultural University,Nanjing 210095,China)  
In order to develop the system for the detection of infectious bursal disease virus(IBDV),nine methods has been carried out for the comparative analysis in the same experiment technique platform.The results indicated that IBDV wild isolates could adapt to the chicken embryo and chicken embryo fibroblast(CEF) cultures by 4-5 passages.The dead embryos presented with inflammation and hemorrhage.Cytopathic effect(CPE) including cell rounding,granulation,reduction,and finally,detachment of cells from the culture surface was observed in virus-infected CEF cultures.Four serotype 1 strains might be further divided into 3 subtypes by cross neutralization test,and the difference of the electrophoresis mobility among the 4 strains could be detected with RT-PCR/SSCP technique.Four methods,including AGP,indirect ELISA,sandwich inhibition ELISA and neutralization test,were simultaneously used to determine the antibodies to IBDV in the different sera,other three methods had a higher sensitivity compared to AGP,with the sensitivity differed by 104 times.The titer of IBDV antigen in the bursal tissue samples could reach up to 1∶10,but in the cell cultures and sick chicken feces didn't present the precipitation line with AGP.In the detection of the above three kind of samples,positive results were obtained with sandwich ELISA,RT-PCR,cDNA probe hybridization,RT-PCR/SSCP and virus isolation methods.
【Fund】: 国家自然科学基金资助项目(30571371);; 江苏省自然科学基金资助项目(BK2003011)
【CateGory Index】: S854.43
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