1株猫嵌杯病毒的分离鉴定及全基因序列分析

doi:10.16431/j.cnki.1671-7236.2023.09.027

Abstract

Abstract: 【Objective】 This study was aimed to isolate Feline calicivirus (FCV) from nasopharynx swab and fecal samples of suspected cats,and provide theoretical basis and reference for studying the biological characteristics and epidemic distribution of FCV.【Method】 11 samples of suspected cats,collected from Jiangmen city,Guangdong province,were screened by RT-PCR.The positive virus was purified by virus plaque purification test and expanded on F81 cells.The purified virus was observed under electron microscope.The median tissue culture infectious dose (TCID₅₀) of the virus was determined by indirect immunofluorescence assay (IFA).The one-step growth curve of the isolate was drawn by measuring the virus titer at different time points of virus collection.The full nucleotide sequence of the isolate was determined,and its genetic evolution and similarity were analyzed.【Result】 A strain of FCV was isolated from faecal disease samples of suspected cats.After purification by plaque,the strain caused the cytopathic effect (CPE) of F81 cells,including shrinkage,rounded,grape cluster aggregation and shedding.Under the electron microscope,the virus appeared as a round,non-envelope virus particle with the diameter of 35-39 nm.The result of IFA showed that the isolated strain could bind specifically with the polyclonal antibody,rabbit anti-FCV VP1-F protein and obvious specific fluorescence could be seen under the fluorescence microscope,and the virus titer was 1×10⁹ TCID₅₀/mL.The whole genome length of the isolate was 7 722 bp,and the nucleotide similarity with FCV-SH strain was the highest (83.94%),belonging to the same branch.VP1 gene had the highest nucleotide similarity with FCV-SH1902 strain (81.32%) and the highest amino acid similarity with FCV-SH1901 strain (89.99%).【Conclusion】 A strain of FCV was successfully isolated and its whole genome sequence was analyzed.This study laid a foundation for the research of pathogenesis and vaccine of FCV.

Key words: Feline calicivirus (FCV); isolation and identification; gene sequencing; genetic evolution analysis

CLC Number:

S852.65^＋9.6

CHEN Changyi, LI Gen, HE Hongling, CHEN Yutong, HUANG Xuezhe, LUO Jun, GUO Xiaofeng. Isolation,Identification and Whole Genome Sequence Analysis of a Strain of Feline Calicivirus[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(9): 3707-3718.

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Isolation,Identification and Whole Genome Sequence Analysis of a Strain of Feline Calicivirus

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