猫1型疱疹病毒分离鉴定及部分生物学特性分析

doi:10.11843/j.issn.0366-6964.2024.07.023

Abstract

Abstract:

This study aimed to obtain feline herpesivirus-1 (FHV-1) isolates from clinical samples through subculture and systematic identification, which laid the foundation for the study of FHV-1 vaccine. Ocular and nasal swabs of suspected FHV-1 infection cats were collected from pet hospitals. Crandell reese feline kidney (CRFK) were used to isolate virus, and evolutionary genetics analysis, ultrastructural observation via electron microscopy and animal regression test were performed on the isolates. The samples were isolated and sub cultured with CRFK cells, followed by identification as FHV-1 by PCR and immunofluorescence, and finally named as "FHV-ZH2202" strain. After the complete gene sequence of the virus was obtained by High-throughput sequencing and gene reference assembly, SNP analysis was conducted between the virus and domestic endemic strains. It was found that the non-synonymous mutant SNPS were mainly distributed in UL22 and US7 genes. Through phylogenetic analysis, FHV-ZH2202 and domestic endemic strains shared high homology in UL22 gene. However, US7 gene harbered by FHV-ZH2202 was with a relatively far relationship with the other domestic endemic strains. The results of animal regression test showed that all cats in infection groups were with typically clinical symptoms such as sneezing, ocular and nasal discharge. However, there was no death case in infection groups. Cats in the FHV-ZH2202 group shed viral DNA in ocular and nasal secretions to from the second day post infection (dpi) to 6-8 days post infection. One FHV-1 virus strain was successfully isolated and part of biological characteristics were analyzed. It was proved that FHV-ZH2202 strain had certain pathogenicity, which laid a foundation for the study of FHV-1 vaccine.

Key words: feline herpesvirus-1, isolation and identification, biological characteristics analysis, high-throughput sequencing, animal regression test

CLC Number:

S852.659.1

Huanqin ZHENG, Xiaomin JIANG, Hong YUE, Baoyan WANG, Yang LIU, Xingxiao ZHANG, Jianlong ZHANG, Hongwei ZHU. Isolation, Identification and Partial Biological Characteristics Analysis of Feline Herpesvirus-1[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(7): 3040-3048.

Figures/Tables 7

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References 17

1	赵涵. 猫疱疹病毒1型宠物猫感染模型的建立[D]. 长春: 吉林农业大学, 2019.
	ZHAO H. Establishment of experimental infection pet cat model by feline herpesvirus type 1[D]. Changchun: Jilin Agricultural University, 2019. (in Chinese)
2	孙春艳,钱鹏,王洁,等.猫疱疹病毒1型的分离鉴定及gD基因序列分析[J].动物医学进展,2020,41(3):30-35.
	SUNC Y,QIANP,WANGJ,et al.Isolation and identification of cat herpesvirus type 1 and sequence analysis of gD gene[J].Progress in Veterinary Medicine,2020,41(3):30-35.
3	LIUC H,LIUY X,QIANP,et al.Molecular and serological investigation of cat viral infectious diseases in China from 2016 to 2019[J].Transbound Emerg Dis,2020,67(6):2329-2335. doi: 10.1111/tbed.13667
4	JASD,FRANCES-DUVERTV,VERNESD,et al.Three-year duration of immunity for feline herpesvirus and calicivirus evaluated in a controlled vaccination-challenge laboratory trial[J].Vet Microbiol,2015,177(1-2):123-131. doi: 10.1016/j.vetmic.2015.03.009
5	牛江婷,伊淑帅,王开,等.猫疱疹病毒1型分子生物学研究进展[J].病毒学报,2017,33(2):274-278.
	NIUJ T,YIS S,WANGK,et al.Advances in research on the molecular biology of feline herpesvirus-1[J].Chinese Journal of Virology,2017,33(2):274-278.
6	HELPSC R,LAITP,DAMHUISA,et al.Factors associated with upper respiratory tract disease caused by feline herpesvirus, feline calicivirus, Chlamydophila felis and Bordetella bronchiseptica in cats: experience from 218 European catteries[J].Vet Rec,2005,156(21):669-673. doi: 10.1136/vr.156.21.669
7	刘美慧,焦翠翠,陈宪平,等.猫疱疹病毒I型核酸可视化检测方法的建立与评价[J].中国兽医学报,2023,43(5):915-923.
	LIUM H,JIAOC C,CHENX P,et al.Establishment and evaluation of visual nucleic acid detection method for feline herpesvirus type I[J].Chinese Journal of Veterinary Science,2023,43(5):915-923.
8	GASKELLR,DAWSONS,RADFORDA,et al.Feline herpesvirus[J].Vet Res,2007,38(2):337-354. doi: 10.1051/vetres:2006063
9	张国军,任洪林,杨举,等.猫疱疹病毒Ⅰ型HB株的分离鉴定及其免疫原性初步评价[J].中国兽医学报,2022,42(5):879-882, 905.
	ZHANGG J,RENH L,YANGJ,et al.Isolation and identification of type 1 HB strain of feline herpesvirus and preliminary evaluation of its immunogenicity[J].Chinese Journal of Veterinary Science,2022,42(5):879-882, 905.
10	于得静. 猫疱疹病毒I型灭活疫苗的制备及宠物猫免疫效果评价[D]. 长春: 吉林农业大学, 2021.
	YU D J. Preparation of feline herpes virus type I inactivated vaccine and evaluation of immune effect in pet cats[D]. Changchun: Jilin Agricultural University, 2021. (in Chinese)
11	SUSSMANM D,MAESR K,KRUGERJ M,et al.A feline herpesvirus-1 recombinant with a deletion in the genes for glycoproteins gI and gE is effective as a vaccine for feline rhinotracheitis[J].Virology,1995,214(1):12-20. doi: 10.1006/viro.1995.9959
12	Mob研究院. 2022年中国宠物消费洞察报告[EB/OL]. [2023-08-09]. https://www.thepaper.cn/newsDetail_forward_20790423.
	Mob Research Institute. 2022 China pet consumption insights report[EB/OL]. [2023-08-09]. https://www.thepaper.cn/newsDetail_forward_20790423. (in Chinese)
13	COHNL A.Feline respiratory disease complex[J].Vet Clin North Am Small Anim Pract,2011,41(6):1273-1289.
14	LAPPINM R,SEBRINGR W,PORTERM,et al.Effects of a single dose of an intranasal feline herpesvirus 1, calicivirus, and panleukopenia vaccine on clinical signs and virus shedding after challenge with virulent feline herpesvirus 1[J].J Feline Med Surg,2006,8(3):158-163.
15	SCOTTF W,GEISSINGERC M.Long-term immunity in cats vaccinated with an inactivated trivalent vaccine[J].Am J Vet Res,1999,60(5):652-658.
16	刘晓晓,刘永相,张继凯,等.猫疱疹病毒1型HR-1毒株的分离鉴定及致病性分析[J].中国兽医学报,2019,39(6):1113-1117.
	LIUX X,LIUY X,ZHANGJ K,et al.Isolation, identification and pathogenicity analysis of feline herpesvirus-1 HR-1 strain[J].Chinese Journal of Veterinary Science,2019,39(6):1113-1117.
17	吴培欣. 2016~2017年广州地区FHV-1的分离鉴定及全基因组测序[D]. 广州: 华南农业大学, 2018.
	WU P X. Separation and identification and whole genome sequencing of feline herpesvirus 1 in Guangzhou from 2016 to 2017[D]. Guangzhou: South China Agricultural University, 2018. (in Chinese)

引物名称 Gene name	引物序列(5′→3′) Primer squence	扩增长度/bp Amplified size	扩增程序 Amplification process
FHV-F	GACGTGGTGAATTATCAGC	288	95 ℃ 3 min; 95 ℃ 15 s, 55 ℃ 15 s, 72 ℃ 30 s, 30 cycles; 72 ℃ 5 min; 4 ℃ ∞
FHV-R	CAACTAGATTTCCACCAGGA
FPV-F	AAAGAGTAGTTGTAAATAATA	681	95 ℃ 3 min; 95 ℃ 30 s, 56 ℃ 30 s, 72 ℃ 40 s, 30 cycles; 72 ℃ 10 min; 4 ℃ ∞
FPV-R	CCTATATCACCAAAGTTAGTAG
FCV-F	GAATTGGCTAARATCTTRCATGA	450	55 ℃ 30 min, 94 ℃ 3 min; 94℃ 30 s, 60 ℃ 30 s, 72 ℃ 30 s, 35 cycles; 72 ℃ 5 min; 4 ℃ ∞
FCV-R	GGRGTTTCAGAGTTDGARGTCA

基因名称 Gene name	流行毒株名称 Strain name	基因突变位点 Gene mutation sites	氨基酸改变 Amino acid mutation sites
UL44	WH2020	C20405T	Glu70Lys
	WH2020	G20879T	Pro204Thr
	GD2018	G20879T	Pro204Thr
	CH-B	G20879T	Pro204Thr
UL22	CH-B	C69213T	Gln512Ser (Gln512*)
		T69454A	Ile593Asn
		T69588A	Cys628Arg
		T70029C	Phe785Leu
US7	WH2020	C122327T	Leu159Phe
		T122346C	Met165Thr
		C122408T	Pro185Ser
	GD2018	C122327T	Leu159Phe
	GD2018	T122346C	Met165Thr
	CH-B	C122327T	Leu159Phe
	CH-B	T122346C	Met165Thr
US8	CH-B	T123211G	Val9Gly
		A123295G	Asp37Gly
		T123780C	Ser199Pro

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Isolation, Identification and Partial Biological Characteristics Analysis of Feline Herpesvirus-1

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