安徽省5株新型鹅星状病毒的分离鉴定与遗传进化分析

doi:10.3969/j.issn.1004-1524.2023.05.08

浙江农业学报 ›› 2023, Vol. 35 ›› Issue (5): 1048-1057.DOI: 10.3969/j.issn.1004-1524.2023.05.08

安徽省5株新型鹅星状病毒的分离鉴定与遗传进化分析

贾北平¹(), 吕炫¹, 杨庆¹, 王一楠¹, 李皖萧¹, 解新迪¹, 朱英奇¹, 王蓓¹, 殷冬冬², 张云凯³, 王晴¹, 王桂军¹^,^*()

1.安徽农业大学动物科技学院,安徽合肥230036
2.安徽省农业科学院畜牧兽医研究所,安徽合肥230001
3.合肥华盟生物技术有限公司,安徽合肥231131

收稿日期:2022-05-28 出版日期:2023-05-25 发布日期:2023-06-01
作者简介:贾北平(1998—),女,安徽六安人,硕士研究生,主要从事畜禽传染病防治研究。E-mail:1853647832@qq.com
通讯作者: *王桂军,E-mail: wgj@ahau.edu.cn
基金资助:
安徽农业大学农业科技推广“揭榜挂帅”专项(2021kjtg001);安徽省家禽产业技术体系岗位专家(AHCYJSTX-06-11);安徽农业大学大学生创新创业训练计划(S202110364035)

Isolation, identification and genetic evolution analysis of novel goose astrovirus in Anhui Province, China

JIA Beiping¹(), LYU Xuan¹, YANG Qing¹, WANG Yinan¹, LI Wanxiao¹, XIE Xindi¹, ZHU Yingqi¹, WANG Bei¹, YIN Dongdong², ZHANG Yunkai³, WANG Qing¹, WANG Guijun¹^,^*()

1. College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China
2. Institute of Animal Husbandry and Veterinary Medicine, Anhui Academy of Agricultural Sciences, Hefei 230001, China
3. Hefei Huameng Biotechnology Co., Ltd., Hefei 231131, China

Received:2022-05-28 Online:2023-05-25 Published:2023-06-01

摘要/Abstract

摘要：

为研究近年来引起雏鹅痛风病的新型鹅星状病毒流行毒株的病原学和遗传学特征,于2018—2021年从安徽省疑似感染新型鹅星状病毒的雏鹅中采集病料,使用健康鹅胚进行病毒分离,并对分离到的毒株进行全基因组测序和序列分析。结果表明,共分离鉴定到5例新型鹅星状病毒感染样品,利用鹅胚盲传3代成功分离到5株新型鹅星状病毒,均可在鹅胚上稳定传代,分别将其命名为AHAU2018、DY-19、ZY02、HR2105/2和HR2110/1株。全基因组测序结果表明,5株分离毒株的基因组全长均为7 175 nt。系统进化树分析结果表明,5株分离毒株均属于致雏鹅痛风的新型鹅星状病毒,但2019年前的毒株与2019年后的毒株处于不同的进化亚分支上。新型鹅星状病毒在我国已发生变异,出现了新的进化亚分支,但优势基因型是否发生变化还有待进一步研究。

关键词: 新型鹅星状病毒, 分离鉴定, 全基因测序, 遗传进化分析

Abstract:

To reveal the etiological and genetic characteristics of the novel goose astrovirus epidemic strains causing goose gout in recent years, the samples were collected from goslings suspected to be infected with the novel goose astrovirus in Anhui Province, China. The viruses were isolated by using healthy goose embryos, and the whole genome sequences of the isolated viruses were determined and analyzed. The results showed that five samples infected with novel goose astrovirus were identified. Five strains of goose astrovirus were successfully isolated from goose embryos by blind transmission for 3 generations. The strains were named as AHAU2018, DY-19, ZY02, HR2105/2 and HR2100/1, respectively. The whole genome sequencing results showed that the total genome length of the isolated strains was 7 175 nt. Phylogenetic tree analysis showed that the five isolated strains belonged to the novel goose astrovirus causing goose gout, and the novel goose astrovirus isolated before 2019 and after 2019 were in different evolutionary sub-branches. The novel goose astrovirus has been mutated in China, and new evolutionary sub branches have emerged. But, whether the dominant genotype has changed remains to be further studied.

Key words: novel goose astrovirus, isolation and identification, whole genome sequencing, genetic evolutionary analysis

中图分类号:

S852.657

贾北平, 吕炫, 杨庆, 王一楠, 李皖萧, 解新迪, 朱英奇, 王蓓, 殷冬冬, 张云凯, 王晴, 王桂军. 安徽省5株新型鹅星状病毒的分离鉴定与遗传进化分析[J]. 浙江农业学报, 2023, 35(5): 1048-1057.

JIA Beiping, LYU Xuan, YANG Qing, WANG Yinan, LI Wanxiao, XIE Xindi, ZHU Yingqi, WANG Bei, YIN Dongdong, ZHANG Yunkai, WANG Qing, WANG Guijun. Isolation, identification and genetic evolution analysis of novel goose astrovirus in Anhui Province, China[J]. Acta Agriculturae Zhejiangensis, 2023, 35(5): 1048-1057.

图/表 10

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编号 No.	分离毒株 Virus strain	临床病例时间 Clinical case date	品种 Variety	送检日龄 Age in days/d	死亡率 Death rate/%
1	AHAU2018	2018年5月 May, 2018	朗德鹅Landes	10	77.8(1 400/1 800)
2	DY19	2019年9月 September, 2019	泰州白鹅Taizhou white	14	23.5(400/1 700)
3	ZY02	2021年4月 April, 2021	皖西白鹅Wanxi white	40	5.0~10.0
4	HR2105/2	2021年5月 May, 2021	朗德鹅Landes	17	40.0(1 000/2 500)
5	HR2110/1	2021年10月 October, 2021	朗德鹅Landes	20	70.4(2 174/3 090)

编号 No.	分离毒株 Virus strain	临床病例时间 Clinical case date	品种 Variety	送检日龄 Age in days/d	死亡率 Death rate/%
1	AHAU2018	2018年5月 May, 2018	朗德鹅Landes	10	77.8(1 400/1 800)
2	DY19	2019年9月 September, 2019	泰州白鹅Taizhou white	14	23.5(400/1 700)
3	ZY02	2021年4月 April, 2021	皖西白鹅Wanxi white	40	5.0~10.0
4	HR2105/2	2021年5月 May, 2021	朗德鹅Landes	17	40.0(1 000/2 500)
5	HR2110/1	2021年10月 October, 2021	朗德鹅Landes	20	70.4(2 174/3 090)

引物名称 Prime name	引物序列 Prime sequences(5'→3')	引物位置 Primer location
GASTV-0-F	AAACAGCGATATGGCGGC	1~18
GASTV-0-R	TGTACGACTTAGTCCAAGGAACG	823~845
GASTV-1-F	CACAGAGGCTACAATGCT	378~395
GASTV-1-R	CCTTCAACAACGACAATGG	1 665~1 683
GASTV-2-F	CAGTCCCTGTACAGATTTTA	1 446~1 465
GASTV-2-R	TCAACTTGTTCATCCTTTAC	2 792~2 811
GASTV-3-F	AGATTGATGAAGCCATTGAG	2 604~2 623
GASTV-3-R	CAGCCCGCCGTTCTGTCTGT	3 939~3 958
GASTV-4-F	AGGCTGTATCAGATATTGAT	3 755~3 774
GASTV-4-R	TCATTTTGTCATTAACGGG	5 014~5 032
GASTV-5-F	TCTGGTGAGTGGCGGACCGA	4 799~4 818
GASTV-5-R	TGTCATAACAGCCCACCAATTGTGT	6 273~6 297
GASTV-6-F	ACAACTGGACAAGGTACC	6 028~6 045
GASTV-6-R	TTTGCGGATTTTAAATGC	7 131~7 148

引物名称 Prime name	引物序列 Prime sequences(5'→3')	引物位置 Primer location
GASTV-0-F	AAACAGCGATATGGCGGC	1~18
GASTV-0-R	TGTACGACTTAGTCCAAGGAACG	823~845
GASTV-1-F	CACAGAGGCTACAATGCT	378~395
GASTV-1-R	CCTTCAACAACGACAATGG	1 665~1 683
GASTV-2-F	CAGTCCCTGTACAGATTTTA	1 446~1 465
GASTV-2-R	TCAACTTGTTCATCCTTTAC	2 792~2 811
GASTV-3-F	AGATTGATGAAGCCATTGAG	2 604~2 623
GASTV-3-R	CAGCCCGCCGTTCTGTCTGT	3 939~3 958
GASTV-4-F	AGGCTGTATCAGATATTGAT	3 755~3 774
GASTV-4-R	TCATTTTGTCATTAACGGG	5 014~5 032
GASTV-5-F	TCTGGTGAGTGGCGGACCGA	4 799~4 818
GASTV-5-R	TGTCATAACAGCCCACCAATTGTGT	6 273~6 297
GASTV-6-F	ACAACTGGACAAGGTACC	6 028~6 045
GASTV-6-R	TTTGCGGATTTTAAATGC	7 131~7 148

分离年份 Year	分离毒株 Virus strain	核苷酸同源性Nucleotide homology/%
		AHAU 2018			DY-19			HR2105/2			HR2110/1			ZY02
		ORF1a	ORF1b	ORF2	ORF1a	ORF1b	ORF2	ORF1a	ORF1b	ORF2	ORF1a	ORF1b	ORF2	ORF1a	ORF1b	ORF2
2016	JSHA	97.6	99.1	98.8	97.7	98.9	98.3	97.5	98.1	97.9	97.3	98.5	98.0	97.5	98.3	97.7
2017	SD01	98.8	99.0	98.5	98.4	98.8	98.1	97.8	98.1	97.7	97.8	98.5	97.8	98.0	98.3	97.4
	GD	97.8	99.0	98.4	98.1	98.8	98.0	97.8	98.0	97.6	97.6	98.4	97.6	97.7	98.2	97.4
	SDPY	98.0	99.0	98.9	98.2	99.0	98.7	97.8	98.2	98.2	97.7	98.6	98.2	97.9	98.4	98.0
2018	AHAU2018	—	—	—	98.1	99.0	98.4	97.7	98.4	98.0	97.6	98.6	98.0	97.8	98.6	97.7
	CXZ	97.8	99.0	99.1	97.8	98.8	98.7	97.5	98.1	98.3	97.3	98.5	98.3	97.6	98.3	98.0
	GTF-04	97.9	98.9	97.8	97.7	98.7	97.6	97.4	98.1	97.1	97.3	98.3	97.3	97.5	98.3	96.9
	GTF-07	97.8	98.6	98.4	97.7	98.4	98.0	97.4	97.6	97.5	97.3	98.0	97.5	97.5	97.8	97.3
	AHAU1	97.8	98.8	98.9	97.9	98.8	98.5	97.5	98.0	98.1	97.4	98.4	98.1	97.5	98.2	97.9
	AHAU4	98.0	98.8	98.9	98.1	98.7	98.5	97.7	97.9	98.1	97.6	98.3	98.1	97.7	98.1	97.8
	XX	98.8	99.4	98.4	98.4	99.2	98.0	98.0	98.4	97.7	97.9	98.8	97.6	98.1	98.6	97.5
2019	DY-19	98.1	99.0	98.4	—	—	—	99.2	98.8	98.3	99.2	99.5	98.9	98.9	99.0	98.1
	SDXT	97.5	98.7	99.0	97.5	98.5	98.7	97.1	97.7	98.2	97.0	98.1	98.2	97.2	97.9	97.9
	HBXG	97.9	98.9	98.8	98.1	98.7	98.4	97.7	97.9	98.0	97.6	98.3	98.2	97.8	98.1	97.8
2020	HNKF-1	97.9	98.8	98.7	99.6	99.7	99.5	99.2	98.6	98.6	99.2	99.5	99.1	98.9	98.8	98.4
	HNSQ-6	98.0	98.9	98.8	99.4	99.7	99.5	99.1	98.7	98.7	99.0	99.4	99.1	98.7	98.9	98.4
2021	HR2105/1	97.7	98.8	98.2	99.2	99.6	98.7	98.7	98.6	98.8	99.4	99.5	98.6	98.8	98.8	98.5
	HR2105/2	97.7	98.4	98.0	98.9	98.8	98.3	—	—	—	98.8	98.5	98.2	99.8	99.8	99.6
	HR2110/1	97.6	98.6	98.0	99.2	99.5	98.9	99.4	98.5	98.2	—	—	—	98.8	98.6	98.0
	ZY02	97.8	98.6	97.7	98.9	99.0	98.1	98.8	99.8	99.6	98.8	98.6	98.0	—	—	—
	ZM	97.9	98.5	98.1	99.0	98.9	98.4	99.0	99.7	99.8	99.0	98.5	98.3	99.8	99.9	99.6

安徽省5株新型鹅星状病毒的分离鉴定与遗传进化分析

Isolation, identification and genetic evolution analysis of novel goose astrovirus in Anhui Province, China

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