动物造模 z-bio 2022-11-18 695

　　OBJECTIVE: To isolate and identify canine parainfluenza virus, analyze the N, HN and F genes of the virus, and study its genetic variation, so as to lay a molecular biology foundation for CPIV diagnosis, treatment and prevention.

　　Methods: The lung tissue of CPIV-positive dogs was inoculated with Vero cells, passed blindly for 3 generations, and the lesions were observed. The 72-hour culture medium was collected for PCR identification, hemagglutination characteristics and morphological observation. At the same time, three pairs of specific primers were used to amplify the entire N, HN and F genes, sequence determination and analysis, and make a phylogenetic tree.

　　RESULTS: A canine parainfluenza virus was successfully isolated from canine lung and named as QF20100726. The virus can agglutinate the blood of guinea pig, pig, chicken and human type O. Electron microscope observation of the virus is round, filamentous and other particles of various shapes and sizes; sequence analysis shows that it is similar to 10 strains of human and dog Compared with the representative parainfluenza virus genes, the nucleotide homology of the N gene is 95.7%~99.8%, and the amino acid homology is 97.4%~99.6%. There are two new amino acid mutations, which are the 257th amino acid. Position changed from V to A, position 301 changed from A to T; HN gene nucleotide homology was 95.5~99.8%, amino acid homology was 96.3~99.6%, F gene nucleotide homology The amino acid homology was 95.6%-99.3%. There are two specific mutations, namely, the 56th position changed from T to S, and the 89th position changed from T to M.

　　Conclusion: The canine parainfluenza virus QF20100726 isolate was successfully isolated, and its hemagglutination properties and ultramicromorphological characteristics were consistent with CPIV characteristics. 1990 (Accession No.: KC237063) is the closest relative; the amino acid of N gene is at position 257 (changed from V to A) and 301 (changed from A to T), and the amino acid of F gene is at position 56 (changed from T to T). became S) and 89 (from T to M) had specific mutations. These data will lay the foundation for the prevention and control of CPIV.