安徽合肥地区2018—2019年犬细小病毒遗传进化分析

doi:10.3969/j.issn.1004-1524.2021.10.04

浙江农业学报 ›› 2021, Vol. 33 ›› Issue (10): 1817-1825.DOI: 10.3969/j.issn.1004-1524.2021.10.04

安徽合肥地区2018—2019年犬细小病毒遗传进化分析

姬凯元(), 邱月阳, 程敖, 蒋书东, 彭梦玲

安徽农业大学安徽省畜禽遗传资源保护与生物育种重点实验室,安徽合肥 230036

收稿日期:2020-09-03 出版日期:2021-10-25 发布日期:2021-11-02
作者简介:姬凯元(1987—),男,山东烟台人,博士,讲师,从事临床兽医学研究。E-mail: 18234487253@163.com
基金资助:
安徽农业大学校青年基金(03087060);安徽省教育厅自然科学基金(KJ2019A0165)

Genetic diversity of Canine parvovirus in Hefei, Anhui Province from 2018 to 2019

JI Kaiyuan(), QIU Yueyang, CHENG Ao, JIANG Shudong, PENG Mengling

Anhui Key Laboratory of Genetic Resources Protection and Biological Breeding for Livestock and Poultry, Anhui Agricultural University, Hefei 230036, China

Received:2020-09-03 Online:2021-10-25 Published:2021-11-02

摘要/Abstract

摘要：

为掌握安徽地区犬细小病毒(CPV)的遗传变异情况,采用PCR法扩增2018—2019年安徽地区收集的10株CPV全基因组,采用生物信息学分析安徽地区CPV遗传进化特点与VP2抗原的变异情况。遗传进化分析发现,安徽地区分离的10株CPV主要以CPV-2c亚型为主,起源于CPV-2aB004(EF011664)和CPV-LZ1(JQ268283)株,正在形成一个独立的分支;NS1基因和VP2基因在遗传进化分支中出现不同步现象,为CPV基因重组提供了依据。VP2蛋白保守性分析发现,安徽分离的CPV毒株的VP2蛋白存在部分氨基酸位点的突变,是否对VP2蛋白的结构和功能产生影响需进一步探究。本研究通过分析安徽地区CPV的遗传进化特点与VP2蛋白氨基酸位点突变特征,为CPV的研究提供了参考。

关键词: 犬细小病毒, 全基因组, 遗传进化

Abstract:

In order to understand the genetic variation of Canine parvovirus (CPV) in Anhui, the whole genome of 10 CPV strains collected in Anhui during 2018—2019 was amplified by PCR, and the genetic evolution characteristics of CPV and the variation of VP2 antigen in Anhui were analyzed by bioinformatics. Genetic evolution analysis showed that the 10 strains of CPV isolated in Anhui were mainly CPV-2c subtypes, originating from CPV-2AB004 (EF011664) and CPV-LZ1 (JQ268283), forming an independent branch. NS1 gene and VP2 gene showed an asynchronous phenomenon in the branch of genetic evolution, which provided the basis for the recombination of CPV gene. Amino acid mutation analysis revealed that VP2 protein of the CPV strain isolated from Anhui Province had some amino acid site mutations, and further research is needed to determine whether it has an impact on the structure and function of VP2 protein.This study provided reference for the study of CPV by analyzing the genetic evolution characteristics of CPV in Anhui Province and the amino acid site mutation characteristics of VP2 protein.

Key words: Canine parvovirus, whole genome, genetic evolution

中图分类号:

S851.3

姬凯元, 邱月阳, 程敖, 蒋书东, 彭梦玲. 安徽合肥地区2018—2019年犬细小病毒遗传进化分析[J]. 浙江农业学报, 2021, 33(10): 1817-1825.

JI Kaiyuan, QIU Yueyang, CHENG Ao, JIANG Shudong, PENG Mengling. Genetic diversity of Canine parvovirus in Hefei, Anhui Province from 2018 to 2019[J]. Acta Agriculturae Zhejiangensis, 2021, 33(10): 1817-1825.

图/表 10

参考文献 27

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引物名称 Primer name	序列 Sequence(5'-3')	位置 Location	产物大小 Product size/bp
CPV-F	CAGGTGATG AATTTGCTACA	3 553-4 583	1 031
CPV-R	AATCTTAAAATAATATGTAATAAAC
Y1-F	ATTCTTTAGAACCAACTGACCAA	1-56	56
Y1-R	CAGCGCGCGTCATCACGTCAT
Y2-F	GCTGCGCGCGCTGCCTAC	53-764	712
Y2-R	TACCGAACAAAGAGTCACCAACCA
C1-F	AAATGGACTGGGAATCGG	490-2 467	1 978
C1-R	GCGGCGTCAGAAGGGTTA
C2-F	AGTCCAACGTGGTCCGAAAT	2 187-3 647	1 461
C2-R	TGGTAAGCCCAATGCTCTAT
C3-F	ACTCCAGCAGCTATGAGA	3 339-4 528	1 118
C3-R	TTTCTAGGTGCTAGTTGAG
U1-F	GCTAAACTAAGAGCCTCTCATACTTGGAATCC	4 392-4 964	573
U1-R	CATAGCGGTCTGGTTGATTAAG CA
U2-F	CTATGCGGTCTGGTTGATTAAGCG	4 960-5 059	100
U2-R	TAGTATCAATCTGTCTTTAAGGGGGG

引物名称 Primer name	序列 Sequence(5'-3')	位置 Location	产物大小 Product size/bp
CPV-F	CAGGTGATG AATTTGCTACA	3 553-4 583	1 031
CPV-R	AATCTTAAAATAATATGTAATAAAC
Y1-F	ATTCTTTAGAACCAACTGACCAA	1-56	56
Y1-R	CAGCGCGCGTCATCACGTCAT
Y2-F	GCTGCGCGCGCTGCCTAC	53-764	712
Y2-R	TACCGAACAAAGAGTCACCAACCA
C1-F	AAATGGACTGGGAATCGG	490-2 467	1 978
C1-R	GCGGCGTCAGAAGGGTTA
C2-F	AGTCCAACGTGGTCCGAAAT	2 187-3 647	1 461
C2-R	TGGTAAGCCCAATGCTCTAT
C3-F	ACTCCAGCAGCTATGAGA	3 339-4 528	1 118
C3-R	TTTCTAGGTGCTAGTTGAG
U1-F	GCTAAACTAAGAGCCTCTCATACTTGGAATCC	4 392-4 964	573
U1-R	CATAGCGGTCTGGTTGATTAAG CA
U2-F	CTATGCGGTCTGGTTGATTAAGCG	4 960-5 059	100
U2-R	TAGTATCAATCTGTCTTTAAGGGGGG

毒株 Strain	登录号 GenBank accession number	亚型 Type	地区 Area	年份 Year
CPV-LZ2	JQ268284	New-CPV-2b	兰州Lanzhou	2011
CPV-GX1	JQ686671	New-CPV-2a	广西Guangxi	2011
CPV-JS2	KF676668	New-CPV-2a	南京Nanjing	2013
CPV-SC02	JX660690	New-CPV-2a	中国China	2011
CPV-s5	KF638400	New-CPV-2a	深圳Shenzhen	2010
CPV-LZ1	JQ268283	New-CPV-2a	兰州Lanzhou	2011
CPV-B004	EF011664	New-CPV-2a	中国China	2004
CPV-OH20219	MF457594	CPV-2c	美国America	2015
CPV-UFMT	KY073269	CPV-2c	巴西Brazil	2015
CPV-UY47	KM457104	CPV-2c	巴西Brazil	2015
CPV-411b.us.98	EU659121	New-CPV-2b	美国Brazil	1988
CPV-447	AY742934	New-CPV-2b	德国Germany	1995
CPV-LaiKa-1993	JN033694	New-CPV-2b	俄罗斯Russia	1993
CPV-Raccoon-349-08	JN867617	New-CPV-2a	美国America	2008
CPV-Raccoon-GA-287-08	JN867615	New-CPV-2a	美国America	2009
CPV-b	M38245	CPV-2	美国America	1988
CPV-5.us.79	EU659116	CPV-2	美国America	1979
CPV-Raccoon-TN-351-09	JN867612	New-CPV-2a	美国America	2009
CPV-Raccoon-VA-118-A-07	JN867610	New-CPV-2a	美国America	2007
CPV-Raccoon-KY-358-B-09	JN867611	New-CPV-2a	美国America	2009
CPV-SC-7	MH476581	CPV-2c	四川Sichuan	2017
CPV-5MGL	MH660909	CPV-2c	内蒙古 Inner Mongolia	2017
CPV-HCM7	LC214969	CPV-2c	越南Vietnam	2013
CPV-SC-9	MF805792	CPV-2c	四川Sichuan	2016
CPV-SC-3	MF805796	CPV-2c	四川Sichuan	2016
CPV-CU21	MH711902	CPV-2c	泰国Thailand	2016
CPV-SH1516	MG013488	CPV-2c	上海Shanghai	2016
CPV-U6	AY742935	CPV-2a	法国France	2016

毒株 Strain	登录号 GenBank accession number	亚型 Type	地区 Area	年份 Year
CPV-LZ2	JQ268284	New-CPV-2b	兰州Lanzhou	2011
CPV-GX1	JQ686671	New-CPV-2a	广西Guangxi	2011
CPV-JS2	KF676668	New-CPV-2a	南京Nanjing	2013
CPV-SC02	JX660690	New-CPV-2a	中国China	2011
CPV-s5	KF638400	New-CPV-2a	深圳Shenzhen	2010
CPV-LZ1	JQ268283	New-CPV-2a	兰州Lanzhou	2011
CPV-B004	EF011664	New-CPV-2a	中国China	2004
CPV-OH20219	MF457594	CPV-2c	美国America	2015
CPV-UFMT	KY073269	CPV-2c	巴西Brazil	2015
CPV-UY47	KM457104	CPV-2c	巴西Brazil	2015
CPV-411b.us.98	EU659121	New-CPV-2b	美国Brazil	1988
CPV-447	AY742934	New-CPV-2b	德国Germany	1995
CPV-LaiKa-1993	JN033694	New-CPV-2b	俄罗斯Russia	1993
CPV-Raccoon-349-08	JN867617	New-CPV-2a	美国America	2008
CPV-Raccoon-GA-287-08	JN867615	New-CPV-2a	美国America	2009
CPV-b	M38245	CPV-2	美国America	1988
CPV-5.us.79	EU659116	CPV-2	美国America	1979
CPV-Raccoon-TN-351-09	JN867612	New-CPV-2a	美国America	2009
CPV-Raccoon-VA-118-A-07	JN867610	New-CPV-2a	美国America	2007
CPV-Raccoon-KY-358-B-09	JN867611	New-CPV-2a	美国America	2009
CPV-SC-7	MH476581	CPV-2c	四川Sichuan	2017
CPV-5MGL	MH660909	CPV-2c	内蒙古 Inner Mongolia	2017
CPV-HCM7	LC214969	CPV-2c	越南Vietnam	2013
CPV-SC-9	MF805792	CPV-2c	四川Sichuan	2016
CPV-SC-3	MF805796	CPV-2c	四川Sichuan	2016
CPV-CU21	MH711902	CPV-2c	泰国Thailand	2016
CPV-SH1516	MG013488	CPV-2c	上海Shanghai	2016
CPV-U6	AY742935	CPV-2a	法国France	2016

病毒Strain	分离地点Area	VP2-297 aa	VP2-426 aa	亚型Type
AHmas 1	马鞍山Maanshan	Ala	Glu	CPV-2c
AHmas 2	马鞍山Maanshan	Ala	Asn	New CPV-2a
AHmas 3	马鞍山Maanshan	Ala	Asp	New CPV-2b
AHmas 4	马鞍山Maanshan	Ala	Glu	CPV-2c
AHmas 6	马鞍山Maanshan	Ala	Glu	CPV-2c
AHmas 9	马鞍山Maanshan	Ala	Glu	CPV-2c
AHmas 11	马鞍山Maanshan	Ala	Glu	CPV-2c
AHmas 12	马鞍山Maanshan	Ala	Asn	New CPV-2a
AHmas 16	马鞍山Maanshan	Ala	Asn	New CPV-2a
AHmas 17	马鞍山Maanshan	Ala	Asn	New CPV-2a
AHmas 18	马鞍山Maanshan	Ala	Glu	CPV-2c
AHhf 1	合肥Hefei	Ala	Glu	CPV-2c
AHhf 2	合肥Hefei	Ala	Glu	CPV-2c
AHhf 5	合肥Hefei	Ala	Glu	CPV-2c
AHhf 6	合肥Hefei	Ala	Glu	CPV-2c
AHhf 7	合肥Hefei	Ala	Glu	CPV-2c
AHhf 13	合肥Hefei	Ala	Glu	CPV-2c
AHhf 15	合肥Hefei	Ala	Glu	CPV-2c
AHhf 19	合肥Hefei	Ala	Glu	CPV-2c
AHhf 20	合肥Hefei	Ala	Glu	CPV-2c
AHhf 21	合肥Hefei	Ala	Glu	CPV-2c
AHhf 22	合肥Hefei	Ala	Glu	CPV-2c
AHhf 23	合肥Hefei	Ala	Glu	CPV-2c
AHhf 24	合肥Hefei	Ala	Glu	CPV-2c
AHhf 27	合肥Hefei	Ala	Glu	CPV-2c
AHhf 28	合肥Hefei	Ala	Glu	CPV-2c
AHcf 1	长丰Changfeng	Ala	Glu	CPV-2c
AHcf 2	长丰Changfeng	Ala	Glu	CPV-2c
AHcf 3	长丰Changfeng	Ala	Glu	CPV-2c
AHcf 4	长丰Changfeng	Ala	Glu	CPV-2c
AHcf 5	长丰Changfeng	Ala	Glu	CPV-2c

安徽合肥地区2018—2019年犬细小病毒遗传进化分析

Genetic diversity of Canine parvovirus in Hefei, Anhui Province from 2018 to 2019

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毒株Strain	3'UTR/bp	NS1/bp	NS2/bp	VP1/bp	VP2/bp	5'UTR/bp
AHmas 3	1-269	270-2 276	270-529	2 386-4 538	2 784-4 538	4 539-5 060
AHmas 17	1-269	270-2 276	270-529	2 386-4 538	2 784-4 538	4 539-5 061
AHhf 1	1-269	270-2 276	270-529	2 386-4 538	2 784-4 538	4 539-5 061
AHmas 9	1-269	270-2 276	270-529	2 386-4 538	2 784-4 538	4 539-5 061
AHcf 4	1-269	270-2 276	270-529	2 386-4 538	2 784-4 538	4 539-5 061
AHmas 2	1-269	270-2 276	270-529	2 386-4 538	2 784-4 538	4 539-5 055
AHhf 7	1-269	270-2 276	270-529	2 386-4 538	2 784-4 538	4 539-5 058
AHhf 27	1-269	270-2 276	270-529	2 386-4 538	2 784-4 538	4 539-5 058
AHhf 28	1-269	270-2 276	270-529	2 386-4 538	2 784-4 538	4 539-5 058
AHmas 16	1-269	270-2 276	270-529	2 387-4 539	2 785-4 539	4 540-5 059

毒株 Strain	分型 Type	氨基酸突变位点Amino acid mutation sites
毒株 Strain	分型 Type	5	13	267	324	370	426	440
AHmas2	New-CPV-2a	Gly	Ser	—	—	Arg	Glu	—
AHmas16	New-CPV-2a	Ala	Pro	—	—	Gln	Asn	—
AHmas17	New-CPV-2a	Ala	Pro	—	—	Gln	Asn	—
CPV-GX1	New-CPV-2a	Ala	Pro	—	—	Gln	Asn	—
CPV-JS2	New-CPV-2a	Ala	Pro	—	—	Gln	Asn	—
CPV-SC02	New-CPV-2a	Ala	Pro	—	—	Gln	Asn	—
CPV-s5	New-CPV-2a	Ala	Pro	—	—	Gln	Asn	—
CPV-LZ1	New-CPV-2a	Ala	Pro	—	—	Gln	Asn	—
CPV-B004	New-CPV-2a	Ala	Pro	—	—	Gln	Asn	—
CPV-Raccoon-349-08	New-CPV-2a	Ala	Pro	—	—	Gln	Asn	—
CPV-Raccoon-GA-287-08	New-CPV-2a	Ala	Pro	—	—	Gln	Asn	—
CPV-Raccoon-TN-351-09	New-CPV-2a	Ala	Pro	—	—	Gln	Asn	—
CPV-Raccoon-VA-118-A-07	New-CPV-2a	Ala	Pro	—	—	Gln	Asn	—
CPV-Raccoon-KY-358-B-09	New-CPV-2a	Ala	Pro	—	—	Gln	Asn	—
AHmas3	New-CPV-2b	—	—	Tyr	Ile	—	—	Ala
CPV-411b.us.98	New-CPV-2b	—	—	Phe	Tyr	—	—	Thr
CPV-447	New-CPV-2b	—	—	Phe	Tyr	—	—	Thr
CPV-LZ2	New-CPV-2b	—	—	Tyr	Ile	—	—	Ala
CPV-LaiKa-1993	New-CPV-2b	—	—	—	—	—	—	—

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