犬副流感病毒,动物造模 z-bio 2021-05-20 672

　　Objective: To isolate and identify canine parainfluenza virus (CPIV), sequence the N, HN and F genes of the virus, study its genetic variation, diagnose, treat and treat CPIV, and provide a molecular basis for prevention.

　　Method: Inject CPIV-positive canine lung tissue into Vero cells, pass them blindly for 3 generations, observe the lesions for PCR identification, observe the coagulation characteristics and morphology, and collect the medium for several hours. At the same time, three pairs of specific primers are used to amplify the entire N, HN and F genes, sequence and analyze them to create a phylogenetic tree.

　　Result: Canine parainfluenza virus was successfully isolated from the lungs of dogs and named it QF20100726. The virus can accumulate type O blood in guinea pigs, pigs, chickens and humans. The electron microscope observes that the virus is round, filamentous, and other particles of various sizes and shapes. Sequence analysis showed that it is similar to 10 human strains. Compared with typical parainfluenza virus genes, the nucleotide homology of the N gene is 95.7% to 99.8%, and the amino acid homology is 97.4% to 99.6%. Two of the amino acids have new mutations, number 257. The position is changed from V to A, and the position 301 is changed from A to T. The nucleotide homology of the HN gene is 95.5-99.8%, the amino acid homology is 96.3-99.6%, the nucleotide homology of the F gene is 94.7%-99.9%, and the amino acid homology is 95.6%-99.3 %. There are two specific mutations. 56 bits have been changed from T to S, and 89 bits have been changed from T to M.

　　Conclusion: Canine parainfluenza virus QF20100726 isolate has been successfully isolated, and its hemagglutination and ultramicroscopic morphological characteristics are consistent with CPIV. , The evolutionary tree of HN and F genes shows that this isolate is the same as the canine parainfluenza virus isolate. 08-1990 (Accession Number: KC237063) This relationship is the closest. The amino acid of the N gene is at position 257 (V to A) and 301 (A to T), and the amino acid of the F gene is at position 56. (From V to A). There is a specific mutation (T becomes S) at position 89 (T to M). These data laid the foundation for the prevention and management of CPIV.