The Research Of Infectious Bursal Disease Virus Reference Materials Used For Nucleic Acids Detection

Infectious bursal disease is an acute,highly contagious viral infection disease which caused by infectious bursal disease virus.This virus is mainly against the chicks’ lymphoid tissue,especially the bursa.Lymphocytes in bursa are destroyed and cannot produce immunoglobulin resulting in reduction of and immune dysfunction.This cause vaccination failure of Marek’s disease,Newcastle disease and at the same time morbidity and mortality increased sharply.So the rapid detection and diagnose of Infectious bursal disease virus play an important role in the prevention and control of it.However the lack of unified reference materials caused many problems in detection and quantification of the virus.In conclusion,the development of nucleic acid reference material really matters in the improvement of accuracy in detection and the guarantee in safety of animal products quality.This research prepares the infectious bursal disease virus nucleic acid reference material using the method of inactivated virus.We have the sterility test and exogenous virus detection on purified virus and the result show that standard material contain no exogenous virus such as AIV,NDV,ALV,GPV,REV,EDSV and so on.We inactivated and initially quantify the IBDV,then blend the lyophilizing protectants and virus,and finally lyophilize the mix.The study establishes the RT-PCR method to detect the content of this reference material,and test the homogeneity and stability of reference material using this method.At the same time we organize other labs to do Cooperative calibration on the reference material,then process statistical analysis on the quantitative data to get the more accurate results with confidence range.The homogeneity research do variance analysis on data and result prove that the reference material is homogeneous for there is no difference within and between the groups.Stability test showed that 7 days(37℃),room temperature(25 degrees),21 days,4 degrees for 4 months and-20 degrees of storage for 1 year did not change significantly.When the confidence rate is 95%,the standard value of the standard material is(2.32±0.67)* 105copies/μL.In summary,a standard of infectious bursal disease virus nucleic acid was prepared and a real-time fluorescence quantitative RT-PCR method of infectious bursal disease virus was established in this study.The reference material has better uniformity and stability,the standard value of the reference material is(2.32±0.67)* 105copies/μL.and can be used to evaluate the laboratory detection methods and selection of detection reagents.