动物造模,综合症病毒多重 z-bio 2022-08-09 586

　　Objective To establish a multiplex PCR detection method for chicken embryo lethal orphan virus (CELO) and chicken egg drop syndrome virus (EDS) and make preliminary applications. Methods Referring to the gene sequences provided by Gen Bank, two pairs of specific primers were designed to amplify the gene sequences of CELO long-fiber protein and EDS hexon protein respectively, and a multiplex PCR method for detecting CELO and EDS was established to investigate the specificity of the method. and sensitivity, and use this method to detect the presence of exogenous avian adenovirus contamination in the influenza vaccine master seed batch. Results The multiplex PCR method successfully amplified two specific target bands, which were verified by sequencing. The specificity of this method is good, and the sensitivity shows that the minimum detection amount of nucleic acid can reach 10-4μg/mL. 12 batches of influenza vaccine master seed batch virus were detected by this method, and the detection results of exogenous avian adenovirus were all negative. Conclusion The multiplex PCR detection method for chicken embryo lethal orphan virus and chicken egg reduction syndrome virus has been successfully established with high sensitivity and specificity. It has high application value and application prospect in the detection of exogenous avian adenovirus from influenza vaccine master seed batch virus