| 犬瘟热病毒SH202003株全基因组序列分析 |
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| 引用本文： | 刘健,白艺兰,鞠厚斌,潘子豪,俞向前,杨显超,朱晓英,葛菲菲,葛杰,于讳茹,陶田谷晟,王建,赵洪进.犬瘟热病毒SH202003株全基因组序列分析[J].中国畜牧兽医,2021,48(7):2559-2568. |
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| 作者姓名： | 刘健 白艺兰 鞠厚斌 潘子豪 俞向前 杨显超 朱晓英 葛菲菲 葛杰 于讳茹 陶田谷晟 王建 赵洪进 |
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| 作者单位： | 1. 上海市动物疫病预防控制中心, 上海 201103;2. 南京农业大学动物医学院, 南京 201195;3. 上海市浦东新区动物疫病预防控制中心, 上海 201299 |
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| 基金项目： | 上海市科技兴农重点攻关项目（沪农科创字[2019]第3-3号）；国家重点研发计划（2017YFD0501702-8） |
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| 摘 要： | 为了解上海地区犬瘟热病毒（Canine distemper virus，CDV）遗传变异情况，本研究采用首尾重叠的11对特异性引物，对CDV上海株SH202003进行RT-PCR扩增，将扩增片段进行反复测序，序列拼接后最终获得了SH202003株全基因组序列，应用Lasergene 7.0和Mega 6.0软件对全基因及H基因进行序列分析，并构建系统进化树。结果显示，SH202003株基因组全长为15 690 bp，编码6种结构蛋白（N、P、M、F、H和L），H和L基因间隔序列为CUA，L和5'端尾随序列为CAA，与Hebei株核苷酸和氨基酸相似性最高，达到98.6%和96.6%，与疫苗株核苷酸相似性在92.2%~94.3%，氨基酸相似性只有82.7%~87.0%；全基因进化树中，SH202003株与流行野毒株在同一分支，与疫苗株在不同的分支；H基因同样与Hebei株亲缘关系最近，核苷酸和氨基酸相似性分别为98.7%和99.5%，与疫苗株Snyder Hill、CDV3、Convac及Onderstepoort亲缘关系较远；SH202003株处于Asia-1型分支，属于Asia-1型强毒株；SH202003株具有9个潜在N-糖基化位点，与强毒株Hebei株一致。研究表明，上海株SH202003属于CDV强毒株，为Asia-1型，其H基因序列相对保守，具有9个潜在N-糖基化位点，但是全基因序列存在较多突变，与疫苗株的匹配度较差，可能是免疫犬依然发生犬瘟热的主要原因。
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| 关 键 词： | 犬瘟热病毒(CDV) 全基因组 序列分析 |
| 收稿时间： | 2021-01-29 |
Genomic Sequencing Analysis of SH202003 Strain of Canine Distemper Virus |
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| LIU Jian,BAI Yilan,JU Houbin,PAN Zihao,YU Xiangqian,YANG Xianchao,ZHU Xiaoying,GE Feifei,GE Jie,YU Huiru,TAO Tiangusheng,WANG Jian,ZHAO Hongjin.Genomic Sequencing Analysis of SH202003 Strain of Canine Distemper Virus[J].China Animal Husbandry & Veterinary Medicine,2021,48(7):2559-2568. |
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| Authors: | LIU Jian BAI Yilan JU Houbin PAN Zihao YU Xiangqian YANG Xianchao ZHU Xiaoying GE Feifei GE Jie YU Huiru TAO Tiangusheng WANG Jian ZHAO Hongjin |
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| Institution: | 1. Shanghai Animal Disease Prevention and Control Center, Shanghai 201103, China;2. College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 201195, China;3. Pudong New Area Animal Disease Control Center in Shanghai, Shanghai 201299, China |
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| Abstract: | To understand the genetic variation of Canine distemper virus (CDV) in Shanghai,the full-length genome of CDV SH202003 strain was sequenced by RT-PCR with 11 pairs of over lapping primers.PCR fragments were obtained and sequenced repeatedly to ensure the mostly possible sequences.The sequenced 11 fragments were spliced to obtain the full-length genomes sequences of CDV SH202003 strain.Alignment analysis of the full-length genome and H gene sequences of SH202003 strain and other CDV sequences published in GenBank was analyzed with Lasergene 7.0 and Mega 6.0 software,and the phylogenetic tree was constructed.The results showed that total length of SH202003 strain was 15 690 bp,encoding 6 structural proteins (N,P,M,F,H and L).The gene interval sequences were CUA between H and L,and CAA between L and 5' trailer,respectively.The SH202003 strain had 98.6% nucleotide sequence similarity and 96.6% amino acid similarity with Hebei strain,and had 92.2% to 94.3% nucleotide sequence similarity and 82.7% to 87.0% amino acid similarity with vaccine strains.Phylogenetic tree analysis showed that SH202003 strain was in the same branch with prevalent virulent strains and distant from the vaccine strains.H gene had 98.7% nucleotide sequence similarity and 99.5% amino acid similarity with Hebei strain,and was distant from the vaccine strains such as Snyder Hill,CDV3,Convac and Onderstepoort.SH202003 strain belonged to Asia-1 subtype (Asia-1 virulent strain).SH202003 strain contained 9 potential N-glycosylation sites,which was consistent with Hebei strain.The results indicated that SH202003 strain was virulent and belonged to Asia-1 type.H gene sequences were relatively conservative and had 9 potential N-glycosylation sites.However,SH202003 strain had many mutations in the full-length genome and low homology with the vaccine strains,which might be the main reason for canine distemper in immunized dogs. |
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| Keywords: | Canine distemper virus (CDV) genomic sequencing analysis |
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