一株鸽副黏病毒Ⅰ型分离鉴定及致病性分析

doi:10.11843/j.issn.0366-6964.2024.09.029

畜牧兽医学报 ›› 2024, Vol. 55 ›› Issue (9): 4051-4060.doi: 10.11843/j.issn.0366-6964.2024.09.029

一株鸽副黏病毒Ⅰ型分离鉴定及致病性分析

张姗¹^,²(), 刘大虎¹^,²^,³, 刘宝京⁴, 梁琳¹^,², 梁瑞英¹^,², 汤新明¹^,², 仇旭升⁵, 丁铲⁵, 丁家波¹^,²^,*(), 侯绍华¹^,*()

1. 中国农业科学院北京畜牧兽医研究所, 北京 100193
2. 农业农村部动物生物安全风险预警及防控重点实验室(北方)/农业农村部兽用生物制品与化学药品重点实验室, 北京 100193
3. 天津农学院动物科学与动物医学学院, 天津市农业动物繁育与健康养殖重点实验室, 天津 300392
4. 北京信和翔科技有限责任公司, 北京 100085
5. 中国农业科学院上海兽医研究所, 上海 200241

收稿日期:2023-11-16 出版日期:2024-09-23 发布日期:2024-09-27
通讯作者: 丁家波,侯绍华 E-mail:zhangshan0276@163.com;dingjiabo@caas.cn;houshaohua@caas.cn
作者简介:张姗(1992-), 女, 河北涞水人, 博士, 主要从事兽医公共卫生研究, E-mail: zhangshan0276@163.com
基金资助:
上海市肉鸽产业技术体系建设(沪农科产字(2023)第12号);中国农业科学院创新工程计划(ASTIP-IAS15)

Isolation, Identification and Pathogenicity Analysis of a Pigeon Paramyxovirus-1 Strain

Shan ZHANG¹^,²(), Dahu LIU¹^,²^,³, Baojing LIU⁴, Lin LIANG¹^,², Ruiying LIANG¹^,², Xinming TANG¹^,², Xusheng QIU⁵, Chan DING⁵, Jiabo DING¹^,²^,*(), Shaohua HOU¹^,*()

1. Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China
2. Key Laboratory of Animal Biosafety Risk Prevention and Control (North)/Key Laboratory of Veterinary Biological Products and Chemicals, Ministry of Agriculture and Rural Affairs, Beijing 100193, China
3. Tianjin Key Laboratory of Agricultural Animal Breeding and Healthy Husbandry, College of Animal Science and Veterinary Medicine, Tianjin Agricultural University, Tianjin 300392, China
4. Bejing Xinhexiang Technology Co., LLC, Beijing 100085, China
5. Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, China

Received:2023-11-16 Online:2024-09-23 Published:2024-09-27
Contact: Jiabo DING, Shaohua HOU E-mail:zhangshan0276@163.com;dingjiabo@caas.cn;houshaohua@caas.cn

摘要/Abstract

摘要：

在天津的病死鸽病料中分离鉴定鸽副黏病毒Ⅰ型(pigeon paramyxovirus-1, PPMV-1), 并对其进行遗传演化、生物学特性、致病性分析, 为鸽副黏病毒Ⅰ型疫苗的研发提供科学依据。采用RT-PCR方法进行病原检测, 在SPF鸡胚上分离纯化病毒; 经遗传演化分析、致病性分析及动物回归试验对病毒分离株进行全面分析。结果显示: 病死鸽的脏器样品经RT-PCR鉴定为NDV核酸阳性; 鸡胚分离纯化后得到1株鸽副黏病毒Ⅰ型毒株, 命名为Pigeon/TJ/CH/020/2020 (TJ20)。通过对病毒F基因测序及进化树分析, 确定该分离毒株属于ClassⅡ类Ⅵ.2.1.1.2.2(原Ⅵk)基因亚型, 裂解位点的氨基酸残基为¹¹²R-R-Q-K-R-F¹¹⁷, 符合NDV强毒株的分子特征。交叉血凝抑制试验显示TJ20株与LaSota疫苗株存在明显的抗原性差异。TJ20株的鸡胚半数感染量(EID₅₀)为10^-8.38·0.1 mL^-1、细胞半数感染量(TCID₅₀)为10^-8.64·0.1 mL^-1、鸡胚平均致死时间(MDT)为62 h、1日龄雏鸡脑内接种致病指数(ICPI)为1.19。动物感染试验结果表明, 1月龄鸽人工感染TJ20株后第4天开始出现嗜睡、垂翅、腹泻、瘫痪、斜颈扭头等新城疫典型临床症状, 发病率和死亡率均为100%。本研究从病死鸽组织中成功分离出一株具有高致病性的Ⅵ.2.1.1.2.2(原Ⅵk)基因亚型的TJ20毒株, 为后续鸽副黏病毒Ⅰ型疫苗研发提供了重要的生物学材料。

关键词: 鸽副黏病毒Ⅰ型, 分离鉴定, Ⅵ.2.1.1.2.2(原Ⅵk)基因亚型, 致病性

Abstract:

A strain of pigeon paramyxovirus-1(PPMV-1) was isolated from the dead pigeon tissue in Tianjin region. To provide scientific basis for the development of PPMV-1 vaccines, the genetic evolution, biological characteristics, and pathogenicity of the virus were analyzed. The pathogens were identified by RT-PCR, virus isolation, purification by chicken embryos, genetic evolution analysis, pathogenicity analysis, and animal regression tests. A strain of PPMV-1 was isolated from the dead pigeons and named Pigeon/TJ/CH/020/2020 (TJ20). Sequence analysis and homology analysis of the virus F gene showed that the strain belonged to sub-genotype Ⅵ.2.1.1.2.2(Ⅵk), and the amino acid residues of the cleavage site were ¹¹²R-R-Q-K-R-F¹¹⁷, which were consistent with the molecular characteristics of NDV virulent strains. Hemagglutination inhibition revealed a noticeable difference between TJ20 and LaSota. The values of TCID₅₀, EID₅₀, MDT and ICPI were 10^-8.64·0.1 mL^-1, 10^-8.38·0.1 mL^-1, 62 h, 1.19. The animal infection experiment found that pigeons showed clinical symptoms on 4 dpc (day 4 post-challenge). The incidence rate and mortality were 100%, and the cloacal detoxification rate could reach 100% on 5 dpc. TJ20 was highly pathogenic to pigeons. This study successfully isolated a highly pathogenic sub-genotype Ⅵ.2.1.1.2.2(Ⅵk) TJ20 strain from samples of dead pigeons, providing important biological materials for the research and development of vaccine of PPMV-1 in the future.

Key words: pigeon paramyxovirus-1, isolation and identification, sub-genotype Ⅵ.2.1.1.2.2(Ⅵk), pathogenicity

中图分类号:

S852.659.5

张姗, 刘大虎, 刘宝京, 梁琳, 梁瑞英, 汤新明, 仇旭升, 丁铲, 丁家波, 侯绍华. 一株鸽副黏病毒Ⅰ型分离鉴定及致病性分析[J]. 畜牧兽医学报, 2024, 55(9): 4051-4060.

Shan ZHANG, Dahu LIU, Baojing LIU, Lin LIANG, Ruiying LIANG, Xinming TANG, Xusheng QIU, Chan DING, Jiabo DING, Shaohua HOU. Isolation, Identification and Pathogenicity Analysis of a Pigeon Paramyxovirus-1 Strain[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(9): 4051-4060.

图/表 8

表 1

图 1

图 2

图 3

图 4

图 5

图 6

表 2

参考文献 35

1	MAYO M A . A summary of taxonomic changes recently approved by ICTV[J]. Arch Virol, 2002, 147 (8): 1655- 1656. doi: 10.1007/s007050200039
2	MILLAR N S , CHAMBERS P , EMMERSON P T . Nucleotide sequence of the fusion and haemagglutinin-neuraminidase glycoprotein genes of Newcastle disease virus, strain Ulster: molecular basis for variations in pathogenicity between strains[J]. J Gen Virol, 1988, 69 (Pt 3): 613- 620.
3	SELIM K M , SELIM A , ARAFA A , et al. Molecular characterization of full fusion protein (F) of Newcastle disease virus genotype VⅡd isolated from Egypt during 2012-2016[J]. Vet World, 2018, 11 (7): 930- 938. doi: 10.14202/vetworld.2018.930-938
4	CZEGLÉDI A , UJVÁRI D , SOMOGYI E , et al. Third genome size category of avian paramyxovirus serotype 1(Newcastle disease virus) and evolutionary implications[J]. Virus Res, 2006, 120 (1-2): 36- 48. doi: 10.1016/j.virusres.2005.11.009
5	GOULD A R , HANSSON E , SELLECK K , et al. Newcastle disease virus fusion and haemagglutinin-neuraminidase gene motifs as markers for viral lineage[J]. Avian Pathol, 2003, 32 (4): 361- 373. doi: 10.1080/0307945031000121112
6	WEI T C , DENG Q M , LI H Q , et al. Molecular characterization of two novel sub-sublineages of pigeon paramyxovirus type 1 in China[J]. Arch Virol, 2018, 163 (11): 2971- 2984. doi: 10.1007/s00705-018-3950-3
7	QIU X S , MENG C C , ZHAN Y , et al. Phylogenetic, antigenic and biological characterization of pigeon paramyxovirus type 1 circulating in China[J]. Virol J, 2017, 14 (1): 186. doi: 10.1186/s12985-017-0857-7
8	LIU X F , WAN H Q , NI X X , et al. Pathotypical and genotypical characterization of strains of Newcastle disease virus isolated from outbreaks in chicken and goose flocks in some regions of China during 1985-2001[J]. Arch Virol, 2003, 148 (7): 1387- 1403. doi: 10.1007/s00705-003-0014-z
9	DIMITROV K M , ABOLNIK C , AFONSO C L , et al. Updated unified phylogenetic classification system and revised nomenclature for Newcastle disease virus[J]. Infect Genet Evol, 2019, 74, 103917. doi: 10.1016/j.meegid.2019.103917
10	ABD EL-HAMID H S , SHAFI M E , ALBAQAMI N M , et al. Sequence analysis and pathogenicity of Avian Orthoavulavirus 1 strains isolated from poultry flocks during 2015-2019[J]. BMC Vet Res, 2020, 16 (1): 253. doi: 10.1186/s12917-020-02470-9
11	HE Y , LU B X , DIMITROV K M , et al. Complete genome sequencing, molecular epidemiological, and pathogenicity analysis of pigeon paramyxoviruses type 1 isolated in Guangxi, China during 2012-2018[J]. Viruses, 2020, 12 (4): 366. doi: 10.3390/v12040366
12	WANG J J , LIU H L , LIU W , et al. Genomic characterizations of six pigeon paramyxovirus type 1 viruses isolated from live bird markets in China during 2011 to 2013[J]. PLoS One, 2015, 10 (4): e0124261. doi: 10.1371/journal.pone.0124261
13	LIU M D , QU Y J , WANG F K , et al. Genotypic and pathotypic characterization of Newcastle disease virus isolated from racing pigeons in China[J]. Poult Sci, 2015, 94 (7): 1476- 1482. doi: 10.3382/ps/pev106
14	韩坤, 梁琳, 李复煌, 等. 鸽新城疫病毒BJ-C分离株基因组测序及系统发育分析[J]. 微生物学通报, 2022, 49 (12): 5034- 5044.
	HAN K , LIANG L , LI F H , et al. Genome sequencing and phylogenetic analysis of Newcastle disease virus BJ-C strain in pigeons[J]. Microbiology China, 2022, 49 (12): 5034- 5044.
15	袁万哲, 邹云婧, 孙继国, 等. 禽流感病毒与新城疫病毒二重RT-PCR检测方法的建立[J]. 河北农业大学学报, 2015, 38 (6): 88- 91.
	YUAN W Z , ZOU Y J , SUN J G , et al. Development of a duplex RT-PCR assay for detection of avian influenza virus and Newcastle disease virus[J]. Journal of Agricultural University of Hebei, 2015, 38 (6): 88- 91.
16	ARCHETTI I , HORSFALL F L Jr . Persistent antigenic variation of influenza A viruses after incomplete neutralization in OVO with heterologous immune serum[J]. J Exp Med, 1950, 92 (5): 441- 462. doi: 10.1084/jem.92.5.441
17	LI Z J , LI Y , CHANG S , et al. Antigenic variation between Newcastle disease viruses of goose and chicken origin[J]. Arch Virol, 2010, 155 (4): 499- 505. doi: 10.1007/s00705-010-0610-7
18	LEI C F , YANG J , HU J , et al. On the calculation of TCID₅₀ for quantitation of virus infectivity[J]. Virol Sin, 2021, 36 (1): 141- 144. doi: 10.1007/s12250-020-00230-5
19	PALDURAI A , KIM S H , NAYAK B , et al. Evaluation of the contributions of individual viral genes to Newcastle disease virus virulence and pathogenesis[J]. J Virol, 2014, 88 (15): 8579- 8596. doi: 10.1128/JVI.00666-14
20	CHEN X , CHEN S Q , CHEN H T , et al. Comparative biology of two genetically closely related Newcastle disease virus strains with strongly contrasting pathogenicity[J]. Vet Microbiol, 2021, 253, 108977. doi: 10.1016/j.vetmic.2020.108977
21	XUE C , XU X H , YIN R F , et al. Identification and pathotypical analysis of a novel Ⅵk sub-genotype Newcastle disease virus obtained from pigeon in China[J]. Virus Res, 2017, 238, 1- 7. doi: 10.1016/j.virusres.2017.05.011
22	LIU H L , WANG Z L , SONG C P , et al. Characterization of pigeon-origin Newcastle disease virus isolated in China[J]. Avian Dis, 2006, 50 (4): 636- 640. doi: 10.1637/7618-042606R1.1
23	杨少华, 崔宁, 张琳, 等. 3株鸽源新城疫病毒的分子特征及对鸽的致病性[J]. 中国兽医学报, 2021, 41 (1): 20- 25.
	YANG S H , CUI N , ZHANG L , et al. Molecular characteristics of three strains of pigeon Newcastle disease virus and its pathogenicity to pigeon[J]. Chinese Journal of Veterinary Science, 2021, 41 (1): 20- 25.
24	罗瑶瑶, 王静静, 王云平, 等. 2014—2017年我国部分地区10株鸽副黏病毒的分布及其分子特征[J]. 中国兽医学报, 2018, 38 (10): 1883-1886, 1931.
	LUO Y Y , WANG J J , WANG Y P , et al. Distribution and molecular characteristics of 10 pigeon paramyxovirus type Ⅰ isolates in some regions of China during 2014 to 2017[J]. Chinese Journal of Veterinary Science, 2018, 38 (10): 1883-1886, 1931.
25	ALEXANDER D J , RUSSELL P H , PARSONS G , et al. Antigenic and biological characterisation of avian paramyxovirus type Ⅰ isolates from pigeons-an international collaborative study[J]. Avian Pathol, 1985, 14 (3): 365- 376. doi: 10.1080/03079458508436238
26	HUTCHISON D C . Paramyxovirus infection in pigeons[J]. Vet Rec, 1984, 115 (13): 335.
27	GANAR K , DAS M , SINHA S , et al. Newcastle disease virus: current status and our understanding[J]. Virus Res, 2014, 184, 71- 81. doi: 10.1016/j.virusres.2014.02.016
28	ALEXANDER D J . Newcastle disease and other avian paramyxoviruses[J]. Rev Sci Tech, 2000, 19 (2): 443- 462.
29	ALDOUS E W , FULLER C M , MYNN J K , et al. A molecular epidemiological investigation of isolates of the variant avian paramyxovirus type 1 virus (PPMV-1) responsible for the 1978 to present panzootic in pigeons[J]. Avian Pathol, 2004, 33 (2): 258- 269.
30	COLLINS M S , STRONG I , ALEXANDER D J . Evaluation of the molecular basis of pathogenicity of the variant Newcastle disease viruses termed "pigeon PMV-1 viruses"[J]. Arch Virol, 1994, 134 (3-4): 403- 411.
31	李仕超, 仇旭升, 刘开春, 等. 六株鸽源新城疫病毒的分离、鉴定及生物信息学分析[J]. 中国动物传染病学报, 2014, 22 (5): 1- 9.
	LI S C , QIU X S , LIU K C , et al. Isolation, identification and bioinformatics analysis of six Newcastle disease virus of pigeon origin[J]. Chinese Journal of Animal Infectious Diseases, 2014, 22 (5): 1- 9.
32	DORTMANS J C F M , KOCH G , ROTTIER P J M , et al. Virulence of pigeon paramyxovirus type 1 does not always correlate with the cleavability of its fusion protein[J]. J Gen Virol, 2009, 90 (Pt 11): 2746- 2750.
33	HEIDEN S , GRUND C , HÖPER D , et al. Pigeon paramyxovirus type 1 variants with polybasic F protein cleavage site but strikingly different pathogenicity[J]. Virus Genes, 2014, 49 (3): 502- 506.
34	钱晶, 刘雪, 王玮, 等. 鸽新城疫病毒Ⅵ-HZ株的分离鉴定及其遗传进化分析[J]. 中国兽医学报, 2022, 42 (4): 651- 656.
	QIAN J , LIU X , WANG W , et al. Isolation, identification and genetic evolution analysis of pigeon Newcastle disease virus Ⅵ-HZ strain[J]. Chinese Journal of Veterinary Science, 2022, 42 (4): 651- 656.
35	裴育, 孙雅丽, 赵烨, 等. 4株鸽新城疫病毒的基因组序列与致病性分析[J]. 病毒学报, 2022, 38 (2): 402- 414.
	PEI Y , SUN Y L , ZHAO Y , et al. Genome sequencing and pathogenicity analyses of four isolates of the Newcastle disease virus in pigeons[J]. Chinese Journal of Virology, 2022, 38 (2): 402- 414.

引物名称Primer name	引物序列(5′→3′) Nucleotide sequence	产物大小/bp Product size
NDV-F	CGAAGAGCCCGTTAGTCAAATC	256
NDV-R	GTATCTTGGCAACCTGGGGAG
ⅥF	GCTTATAGTCAGTTTACCTGCC	1 931
ⅥR	GTGGTAGAACAGATGTTGTGAAG

组别Group	毒株Strain	数量/只Number	口咽Oropharyngeal				泄殖腔Cloacal
组别Group	毒株Strain	数量/只Number	第3天3 dpc	第5天5 dpc	第7天7 dpc	第10天10 dpc	第3天3 dpc	第5天5 dpc	第7天7 dpc	第10天10 dpc
感染组Infection group	TJ20	10	0/10	3/10	3/9	2/3	1/10	10/10	9/9	3/3
对照组Control group	/	10	0/10	0/10	0/10	0/10	0/10	0/10	0/10	0/10

[1]	谢碧林, 林志敏, 林彬彬, 徐以娟, 林锋强, 闫露, 伍慧妮, 李翠婷, 周海欧, 李兆龙. 鸭疫里氏杆菌LC1和CX1株的分离鉴定及致病性分析[J]. 畜牧兽医学报, 2024, 55(9): 4196-4203.
[2]	王佳宁, 张自强, 王帅帅, 刘玉梅. 兔源大肠杆菌、奇异变形杆菌和球虫混合感染及药物敏感性分析[J]. 畜牧兽医学报, 2024, 55(9): 4204-4212.
[3]	王艳, 高亚东, 蒋成辉, 曾巧英. 一株鹅源禽腺病毒4型的分离及致病性[J]. 畜牧兽医学报, 2024, 55(9): 4232-4240.
[4]	张帆帆, 李杰茂, 谭佳, 黄江南, 吴玲, 韦启鹏, 康昭风. 禽偏肺病毒的研究进展[J]. 畜牧兽医学报, 2024, 55(8): 3344-3353.
[5]	杨程, 刘野, 程宁, 王凯月, 李欣蕾, 孙久英, 韩俊平, 李文军, 王欢欢, 邵笑, 程雪娇, 孙英峰. 一株PRRSV-2谱系1.8与1.5重组毒株的基因组特征分析[J]. 畜牧兽医学报, 2024, 55(8): 3570-3578.
[6]	程玉婷, 阿比克哈莫, 杨晨, 张丁中, 任云鑫, 岳华, 汤承. 山羊Aichivirus C的分离鉴定及演化分析[J]. 畜牧兽医学报, 2024, 55(8): 3612-3622.
[7]	孙雨点, 宋紫玥, 张洪亮, 秦志华, 单虎, 杨瑞梅. 鸭短喙与侏儒综合征病毒分离与鉴定[J]. 畜牧兽医学报, 2024, 55(8): 3623-3630.
[8]	王奇林, 曹润来, 王威阳, 张博, 刘志杰, 王晓旭. 狐狸流产胎儿体内肺炎克雷伯菌的分离鉴定及耐药性分析[J]. 畜牧兽医学报, 2024, 55(8): 3640-3648.
[9]	郑焕琴, 姜晓敏, 岳红, 王宝岩, 刘洋, 张兴晓, 张建龙, 朱洪伟. 猫1型疱疹病毒分离鉴定及部分生物学特性分析[J]. 畜牧兽医学报, 2024, 55(7): 3040-3048.
[10]	刘维哲, 罗成刚, 袁蓉, 廖艺杰, 文艺悯, 孙莹, 俞恩波, 曹三杰, 黄小波. 一株猪流行性腹泻病毒强毒株的分离与鉴定[J]. 畜牧兽医学报, 2024, 55(7): 3049-3063.
[11]	李继桐, 朱彤, 吕俊峰, 高月花, 胡峰, 于可响, 宋敏训, 王建琳, 李玉峰. 三株新型鸭源微RNA病毒分离毒株的全基因组序列分析[J]. 畜牧兽医学报, 2024, 55(7): 3075-3084.
[12]	班玛王清, 陈曦, 岳怡, 苏玉蓉, 岳华, 汤承. 一株牛呼吸道冠状病毒的分离鉴定及部分生物学特征[J]. 畜牧兽医学报, 2024, 55(7): 3094-3104.
[13]	刘博华, 符汉宇, 王玉恒, 索朗斯珠, 牛家强, 包玉花, 李家奎, 徐业芬. 西藏那曲市牦牛源B型多杀性巴氏杆菌的分离鉴定及基因组分析[J]. 畜牧兽医学报, 2024, 55(7): 3105-3118.
[14]	杨坤, 马静文, 周新瑞, 罗烈柱, 刘喆, 胡自强, 武星辰, 梁立滨, 高诗敏. 三株传染性法氏囊病病毒重组毒株的致病性研究[J]. 畜牧兽医学报, 2024, 55(6): 2550-2559.
[15]	周宁, 汤承, 许佳, 岳华, 陈曦. 猫泛白细胞减少症病毒A91S变异株对猫的致病性及基因组特征研究[J]. 畜牧兽医学报, 2024, 55(6): 2560-2568.

一株鸽副黏病毒Ⅰ型分离鉴定及致病性分析

Isolation, Identification and Pathogenicity Analysis of a Pigeon Paramyxovirus-1 Strain

PDF

可视化

摘要/Abstract

引用本文

使用本文

图/表 8

参考文献 35

相关文章 15

编辑推荐

Metrics

本文评价