Acta Agriculturae Zhejiangensis ›› 2021, Vol. 33 ›› Issue (10): 1817-1825.DOI: 10.3969/j.issn.1004-1524.2021.10.04
• Animal Science • Previous Articles Next Articles
JI Kaiyuan(
), QIU Yueyang, CHENG Ao, JIANG Shudong, PENG Mengling
Received:2020-09-03
Online:2021-10-25
Published:2021-11-02
CLC Number:
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.
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URL: http://www.zjnyxb.cn/EN/10.3969/j.issn.1004-1524.2021.10.04
| 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 |
Table 2 Primers for identification and complete genome sequence amplification
| 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 |
Table 2 Reference sequences of CPV-2 strains
| 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 |
Table 3 Key amino acid loci of CPV strains in Anhui
| 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 |
| 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 |
Table 4 Genome structure of 10 strains CPV Anhui isolated
| 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 | ||||||
|---|---|---|---|---|---|---|---|---|
| 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 | — | — | — | — | — | — | — |
Table 5 Statistical results of VP2 protein amino acid mutation sites
| 毒株 Strain | 分型 Type | 氨基酸突变位点Amino acid mutation sites | ||||||
|---|---|---|---|---|---|---|---|---|
| 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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