4份临床样品中羊口疮病毒F1L、B2L和GIF基因的进化分析

林裕胜; 刘维巍; 张龙; 江锦秀; 张靖鹏; 刘庆华; 胡奇林

doi:10.19303/j.issn.1008-0384.2022.06.009

4份临床样品中羊口疮病毒F1L、B2L和GIF基因的进化分析

doi: 10.19303/j.issn.1008-0384.2022.06.009

林裕胜^1,,
刘维巍^{1, 2,},
张龙^{1, 2},
江锦秀¹,
张靖鹏¹,
刘庆华²,
胡奇林^1, ,

1.
福建省农业科学院畜牧兽医研究所，福建　福州　350013
2.
福建农林大学动物科学学院，福建　福州　350002

基金项目: 福建省农业高质量发展超越“5511”协同创新工程项目（XTCXGC2021008）；福建省科技计划公益类专项（2021R1026007）；福建省自然科学基金项目（2021J01484）；福建省农业科学院科技创新团队建设项目（CXTD2021007-2）

详细信息

作者简介:
林裕胜（1988−），男，硕士，助理研究员，主要从事动物传染病研究（E-mail：369807285@qq.com）

刘维巍（1999−），男，硕士，主要从事动物传染病研究（E-mail：2505155891@qq.com）

通讯作者:
胡奇林（1963-），男，硕士，研究员，主要从事动物传染病学和免疫学研究（E-mail：hql562713@163.com）

中图分类号: S 852.65
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出版历程
- 收稿日期: 2022-03-31
- 录用日期: 2022-03-31
- 修回日期: 2022-04-14
- 网络出版日期: 2022-06-20
- 刊出日期: 2022-06-28

Genetic Evolution of F1L, B2L, and GIF of Orf Virus in Four Clinical Samples

LIN Yusheng^1
,,
LIU Weiwei^{1, 2
,},
ZHANG Long^{1, 2},
JIANG Jinxiu¹,
ZHANG Jingpeng¹,
LIU Qinghua²,
HU Qilin^{1
, ,}

1.
Institute of Animal Husbandry & Veterinary Medicine, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian　350013, China
2.
College of Animal Science, Fujian Agriculture and Forestry University, Fuzhou, Fujian　350002, China

摘要

摘要: 目的研究2021年福建省部分羊场羊口疮病毒（Orf virus, ORFV）分子流行病学情况。方法对2021年采集的4份病料经PCR鉴定ORFV阳性的临床样品分别克隆其F1L、B2L和GIF基因，将获得的克隆质粒送往测序公司进行测序，将测序拼接后的序列上传至GenBank数据库中，利用生物信息学软件进行对比分析。结果 4份临床样品中ORFV的F1L、B2L和GIF基因核苷酸序列相似性分别为98.0%～98.8%、98.4%～99.5%和97.2%～99.6%；与国内流行毒株核苷酸序列相似性分别为97.4%～99.4%、97.4%～99.9%和95.7%～99.0%；与国外流行毒株核苷酸序列相似性分别为97.5%～98.6%、97.9%～98.9%和95.6%～98.0%；与德国D1701弱毒株核苷酸序列相似性分别为96.5%～96.8%、98.0%～98.7%和96.0%～96.4%，与NZ2参考株核苷酸序列相似性分别为97.7%～98.1%、97.5%～98.1%和95.9%～96.1%。遗传进化树结果显示4份临床样品中ORFV与吉林（NA17、SY17和CL18株）、福建SJ1株和印度MP株亲缘关系较近；与美国OV-IA82株、德国D1701弱毒株、新西兰NZ2参考株和中国疫苗株亲缘关系较远；表明当前流行毒株可能是本省毒株、国内吉林GZ18株和国外印度MP株的重组毒株。结论获得4份临床样品中ORFV B2L、F1L和GIF基因序列并上传至GenBank数据库中，丰富了ORFV相关基因序列的数据库，同时阐明福建省部分羊场当前ORFV的流行特征，为福建省今后有效防控Orf提供理论依据。
- 羊口疮病毒 /
- F1L基因 /
- B2L基因 /
- GIF基因 /
- 进化分析
Abstract: Objective Molecular epidemiology of the Orf virus (ORFV) in the diseased sheep and goats from farms in Fujian Province in 2021 were studied. Method The ORFV genes, F1L, B2L, and GIF, from 4 clinical samples of the diseased animals were confirmed positive by PCR prior to cloning for sequence determination. The data were uploaded to GenBank for a comparative analysis using bioinformatics software. Result The nucleotide sequence similarities were 98.0%–98.8% on F1L , 98.4%–99.5% on B2L, and 97.2%–99.6% on GIF; those related to the domestic epidemic strains, 97.4%–99.4% on F1L, 97.4%–99.9% on B2L, and 95.7%–99.0% on G1F; those related to the foreign epidemic strains, 97.5%–98.6% on F1L, 97.9%–98.9% on B2L, and 95.6%–98.0% on G1F; those related to the German D1701 attenuated strain, 96.5%–96.8% on F1L, 98.0%–98.7% on B2L, and 96.0%–96.4% on G1F; and those related to the NZ2 reference strain, 97.7%–98.1% on F1L, 97.5%–98.1% on B2L, and 95.9%–96.1% on G1F. The phylogenetic tree showed the ORFVs closely related to the strains from Jilin (NA17, SY17, and CL18), Fujian (SJ1), and India (MP), but remotely with OV-IA82 from the USA, D1701 attenuated strain from German, NZ2 reference strain from New Zealand, and the vaccine strain from China. It appeared that the current epidemic strain might be a recombination of Fujian, Jilin GZ18, and Indian MP strains. Conclusion The sequences of B2L, F1L, and GIF from the 2021 clinical specimens were uploaded to GenBank database enriching its collection. This study characterized the ORFVs found in the sheep and goats infected by the epidemic and facilitated further studies in the disease control and prevention.
- Orf virus /
- F1L /
- B2L /
- GIF /
- evolution analysis

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图 1 F1L基因的遗传进化树

Figure 1. Phylogenetic tree analysis on F1L

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图 2 B2L基因的遗传进化树

Figure 2. Phylogenetic tree analysis on B2L

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图 3 GIF基因的遗传进化树

Figure 3. Phylogenetic tree analysis on GIF

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表 1 引物序列

Table 1. Sequences of primers

引物名称 Primers	引物序列 Sequences（5'-3'）	退火温度 Annealing/℃	片段大小 Product size/bp
F1L-F	CGGAATTCGATGGATCCACCCGAAATCACG	55	1029
F1L-R	CCGTCGAGTCACACGATGGCCGTGACCA	55	1029
B2L-F	CGGAATTCGATGTGGCCGTTCTCCTCCATC	55	1137
B2L-R	CCGTCGAGTTAATTTATTGGCTTGCAGAACTCC	55	1137
GIF-F	CGGAATTCGATGGCGTGCCTCAGAGTGTT	52	798
GIF-R	CCGTCGAGTTAGCGCGCCGTCTGCATGC	52	798

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表 2 ORFV 参考毒株

Table 2. Reference ORFV strains

毒株 Virus	登录号 Locus	分离地 Isolate	时间 Time
NZ2 OV-SA00 OV-IA82 D1701 GZ18 SJ1 NP NA17 YX GO OV-HN3/12 NA1/11 MP SY17 CL18 TVL China vaccine	DQ184476 AY386364 AY386363 HM133903 MN648218 KP010356 KP010355 MG674916 KP010353 KP010354 KY053526 KF234407 MT332357 MG712417 MN648219 MN454854 JQ904789	New Zealand USA USA Germany Jilin, China Fujian, China Fujian, China Jilin, China Fujian, China Fujian, China Guangzhou, China Guangzhou, China India Jilin, China Jilin, China West Texas, USA Gansu, China	2005 2003 2003 2010 2018 2015 2011 2016 2012 2012 2011 2013 2017 2016 2018 2019 2012

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