一株猫泛白细胞减少症病毒的分离鉴定及其VP2和NS1基因的变异分析

doi:10.11843/j.issn.0366-6964.2019.05.022

畜牧兽医学报 ›› 2019, Vol. 50 ›› Issue (5): 1106-1112.doi: 10.11843/j.issn.0366-6964.2019.05.022

• 研究简报 • 上一篇

一株猫泛白细胞减少症病毒的分离鉴定及其VP2和NS1基因的变异分析

刘琪^1,2, 史利军^1,2, 梁琳^1,2, 张玲玲^1,2, 袁维峰^1,2, 梁瑞英^1,2, 李金祥^1,2*, 崔尚金^1,2*

1. 中国农业科学院北京畜牧兽医研究所, 北京 100193;
2. 农业部兽用药物与诊断技术北京科学观测实验站, 北京 100193

收稿日期:2018-08-22 出版日期:2019-05-23 发布日期:2019-05-23
通讯作者: 李金祥,主要从事动物病原学与流行病学研究,E-mail:lijinxiang@caas.cn;崔尚金,主要从事动物病原学与流行病学研究,E-mail:cuishangjin@caas.cn
作者简介:刘琪(1994-),女,辽宁朝阳人,硕士生,主要从事动物病原学与流行病学研究,E-mail:1834820417@qq.com
基金资助:
“十三五”国家重点研发计划（2016YFD0501003）；中国农业科学院创新工程项目（ASTIP-IAS15）

Isolation and Identification of a Feline Panleukopenia Virus and Genetic Variation Analysis of Its VP2 and NS1

LIU Qi^1,2, SHI Lijun^1,2, LIANG Lin^1,2, ZHANG Lingling^1,2, YUAN Weifeng^1,2, LIANG Ruiying^1,2, LI Jinxiang^1,2*, CUI Shangjin^1,2*

1. Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China;
2. Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Technology of Beijing, Ministry of Agriculture, Beijing 100193, China

Received:2018-08-22 Online:2019-05-23 Published:2019-05-23

摘要/Abstract

摘要：

为分析当前猫泛白细胞减少症病毒（FPLV）基因组遗传变异情况，对FPLV胶体金试纸条检测结果为阳性的猫粪进行病毒分离，通过血凝、分子生物学等试验对分离株进行鉴定。结果表明：病毒在CRFK细胞上盲传5代后产生明显细胞病变，电镜观察病毒粒子直径约25 nm，分离病毒的血凝效价为1：256，将分离株命名为FPLV BJ04。基因组扩增结果显示，FPLV BJ04基因组大小为5 118 bp。决定病毒宿主范围和抗原性的VP2蛋白系统进化树分析表明，FPLV BJ04株与2017年意大利分离株KX434462株位于同一分支；分离株的VP2、NS1基因与GenBank收录的FPLV VP2、NS1基因相似性分别为99.1%~99.5%和99.0%~99.5%。动物回归试验中攻毒组猫出现临床症状和病理变化。本研究对分离的FPLV北京流行株的VP2和NS1基因进行遗传变异分析，可为更好地预防和控制FPLV提供基础。

Abstract:

To analyze genetic variation of feline panleukopenia virus (FPLV), in this study, a fecal sample was collected from a cat infected FPLV in Beijing and was used to isolate FPLV by CRFK cells, and identified by hemagglutination assay (HA), molecular biology. The results showed that cytopathic effects could be observed in CRFK cells. The diameter of virus particle about 25 nm was observed by electron microscope. The HA blood coagulation rate was 1:256. The full genome was cloned, sequenced and analyzed. The results showed that the isolated virus had a genome of 5 118 bp, phylogenetic tree of VP2 showed that FPLV BJ04 strain and KX434462 strain were on the same small branch. The isolated virus, according to VP2 gene homology analysis, shared from 99.1% to 99.5% homology with reference strains, and NS1 gene shared from 99.0% to 99.5%. Animal regression test results also suggested that clinical symptoms and pathological changes were observed in the challenge group. In this study, an epidemic strain of FPLV in Beijing was successfully isolated, and the genetic variation of VP2 and NS1 gene were analyzed. The results provide basic information for the better prevention and control of FPLV.

中图分类号:

S852.659.2

刘琪, 史利军, 梁琳, 张玲玲, 袁维峰, 梁瑞英, 李金祥, 崔尚金. 一株猫泛白细胞减少症病毒的分离鉴定及其VP2和NS1基因的变异分析[J]. 畜牧兽医学报, 2019, 50(5): 1106-1112.

LIU Qi, SHI Lijun, LIANG Lin, ZHANG Lingling, YUAN Weifeng, LIANG Ruiying, LI Jinxiang, CUI Shangjin. Isolation and Identification of a Feline Panleukopenia Virus and Genetic Variation Analysis of Its VP2 and NS1[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(5): 1106-1112.

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参考文献

[1] STEINEL A, MUNSON L, VAN VUUREN M, et al. Genetic characterization of feline parvovirus sequences from various carnivores[J]. J Gen Virol, 2000, 81(2):345-350.
[2] 李刚,蔡宝祥,张振兴. 猫泛白细胞减少症病毒的分离与鉴定[J]. 病毒学报, 1985, 1(4):349-353.
LI G, CAI B X, ZHANG Z X. Isolation and identification of feline panleukopenia virus[J]. Chinese Journal of Virology, 1985, 1(4):349-353. (in Chinese)
[3] JUNGE R E, BAUMAN K, KING M, et al. A serologic assessment of exposure to viral pathogens and Leptospira in an urban raccoon (Procyon lotor) population inhabiting a large zoological park[J]. J Zoo Wild Med, 2007, 38(1):18-26.
[4] 黄翠琴, 范克伟,余佳,等. 华南虎疑似感染猫细小病毒的PCR鉴定与病理组织学检查[J]. 中国动物传染病学报, 2017, 25(6):43-47.
HUANG C Q, FAN K W, YU J, et al. A case of feline parvovirus virus infection of a south Chinatiger[J]. Chinese Journal of Animal Infectious Diseases, 2017, 25(6):43-47. (in Chinese)
[5] 饶家辉, 王玉平,雷连成. 猫细小病毒、犬细小病毒、貂细小病毒的特征比较[J]. 中国畜牧兽医, 2009, 36(7):166-168.
RAO J H, WANG Y P, LEI L C. Characteristic comparison of feline panleukopenia virus, canine parvovirus and mink enteritis virus[J]. China Animal Husbandry & Veterinary Medicine, 2009, 36(7):166-168. (in Chinese)
[6] GOVINDASAMY L, HUEFFER K, PARRISH C R, et al. Structures of host range-controlling regions of the capsids of canine and feline parvoviruses and mutants[J]. J Virol, 2003, 77(22):12211-12221.
[7] LÍPEZ DE TURISO J A, CORTÉS E, MARTÍNEZ C, et al. Recombinant vaccine for canine parvovirus in dogs[J]. J Virol, 1992, 66(5):2748-2753.
[8] 殷震,刘景华. 动物病毒学[M]. 2版. 北京:科学出版社, 1997:1145-1173.
YIN Z, LIU J H. Animalvirology[M]. 2nd ed. Beijing:Science Press, 1997:1145-1173. (in Chinese)
[9] 刘维全, 范泉水,江禹,等. 肉食兽细小病毒通用PCR诊断技术的建立[J]. 中国兽医学报, 2001, 21(3):249-251.
LIU W Q, FAN Q S, JIANG Y, et al.Establishment of a commonly used PCR technique for detection of Carnivore parvoviruses[J]. Chinese Journal of Veterinary Science, 2001, 21(3):249-251. (in Chinese)
[10] 柏丽华. 北京地区犬细小病毒病的分子流行病学调查研究[D]. 北京:中国农业科学院, 2011.
BAI L H. The molecular epidemiology investigation of Canine parvovirus infection in Beijing[D]. Beijing:Chinese Academy of Agricultural Sciences, 2011. (in Chinese)
[11] 田丽红, 华育平. 虎源猫泛白细胞减少症病毒VP2基因的原核表达及其抗原特性[J]. 中国兽医学报, 2010, 30(5):602-606.
TIAN L H, HUA Y P. Prokaryotic expression of feline panleukopenia virus(VP2) isolated from tiger and its antigenic characteristics[J]. Chinese Journal of Veterinary Science, 2010, 30(5):602-606. (in Chinese)
[12] 程楠,赵永坤,于永乐,等. 猫细小病毒HH-1/86株理化性质鉴定及全基因组测序[J]. 中国兽医学报, 2017, 37(5):818-822.
CHENG N, ZHAO Y K, YU Y L, et al. The physical and chemical characterization of FPV strain HH-1/86 and its whole genome sequenceanalysis[J]. Chinese Journal of Veterinary Science, 2017, 37(5):818-822. (in Chinese)
[13] TRUYEN U, AGBANDJE M, PARRISH C R. Characterization of the feline host range and a specific epitope of feline panleukopenia virus[J]. Virology, 1994, 200(2):494-503.
[14] DECARO N, DESARIO C, MICCOLUPO A, et al. Genetic analysis of feline panleukopenia viruses from cats with gastroenteritis[J]. Gen Virol, 2008, 89(9):2290-2298.
[15] 卞赛赛, 梁琳,高窦,等. 水貂肠炎病毒VP2基因的原核表达及多克隆抗体的制备[J]. 中国兽医科学, 2017, 47(5):597-602.
BIAN S S, LIANG L, GAO D, et al. Prokaryotic expression of VP2 gene of mink enteritis virus and preparation of anti-serum[J]. Chinese Veterinary Science, 2017, 47(5):597-602. (in Chinese)
[16] STUCKER K M, PAGAN I, CIFUENTE J O, et al. The role of evolutionary intermediates in the host adaptation of canine parvovirus[J]. J Virol, 2012, 86(3):1514-1521.
[17] YANG S T, WANG S J, FENG H, et al. Isolation and characterization of feline panleukopenia virus from a diarrheic monkey[J]. Vet Microbiol, 2010, 143(2-4):155-159.
[18] 饶家辉, 王玉平,雷连成,等. 猫细小病毒JL1-05株的分离与鉴定[J]. 中国兽医学报, 2009, 29(6):716-720.
RAO J H, WANG Y P, LEI L C, et al. Isolation and identification of feline panleukopenia virus strain JL1-05[J]. Chinese Journal of Veterinary Science, 2009, 29(6):716-720. (in Chinese)
[19] HOELZER K, SHACKELTON L A, PARRISH C R, et al. Phylogenetic analysis reveals the emergence, evolution and dispersal of carnivore parvoviruses[J]. J Gen Virol, 2008, 89(9):2280-2289.

一株猫泛白细胞减少症病毒的分离鉴定及其VP2和NS1基因的变异分析

Isolation and Identification of a Feline Panleukopenia Virus and Genetic Variation Analysis of Its VP2 and NS1

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