2021—2022年我国部分地区猪繁殖与呼吸综合征病毒ORF5基因变异分析

doi:10.11843/j.issn.0366-6964.2023.09.021

畜牧兽医学报 ›› 2023, Vol. 54 ›› Issue (9): 3812-3823.doi: 10.11843/j.issn.0366-6964.2023.09.021

2021—2022年我国部分地区猪繁殖与呼吸综合征病毒ORF5基因变异分析

王志远^1,2,3, 刘博奇⁴, 许志颖^1,2, 徐思佳^1,2, 邢家宝^1,2, 张桂红^2,3, 王衡^1,2*, 孙彦阔^1,2,3*

1. 华南农业大学兽医学院, 广东省临床重大疾病综合防控重点实验室, 广州 510642;
2. 岭南现代农业科学与技术广东省实验室茂名分中心, 茂名 525000;
3. 华南农业大学国家生猪种业工程技术研究中心, 广州 510642;
4. 广东海大畜牧兽医研究院有限公司, 广州 511400

收稿日期:2022-10-26 发布日期:2023-09-22
通讯作者: 孙彦阔,主要从事兽医传染病研究,E-mail:yankuosun@scau.edu.cn;王衡,主要从事兽医病理学研究,E-mail:wangheng2009@scau.edu.cn
作者简介:王志远(1996-),男,山东招远人,硕士生,主要从事兽医病理学及兽医病毒分子生物学研究,E-mail:978419898@qq.com;刘博奇(1982-),女,助理研究员,硕士,主要从事动物病理与免疫学研究,E-mail:liuboqi0716@126.com
基金资助:
国家自然科学基金项目(32102704);茂名实验室科研启动项目(2021TDQD002);财政部和农业农村部:国家现代农业产业技术体系

Analysis of ORF5 Gene Variation of Porcine Reproductive and Respiratory Syndrome Virus in Local Regions of China from 2021 to 2022

WANG Zhiyuan^1,2,3, LIU Boqi⁴, XU Zhiying^1,2, XU Sijia^1,2, XING Jiabao^1,2, ZHANG Guihong^2,3, WANG Heng^1,2*, SUN Yankuo^1,2,3*

1. Key Laboratory of Comprehensive Prevention and Control for Severe Clinical Animal Diseases of Guangdong Province, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China;
2. Maoming Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Maoming 525000, China;
3. National Engineering Research Center for Breeding Swine Industry, South China Agricultural University, Guangzhou 510642, China;
4. Guangdong Haid Animal Husbandry and Veterinary Research Institute Co. Ltd, Guangzhou 511400, China

Received:2022-10-26 Published:2023-09-22

摘要/Abstract

摘要： 为深入了解我国猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndrome virus, PRRSV)的流行和遗传变异情况,2021-2022年,本课题组在我国13个省份约50个养殖场收集了117份临床疑似猪繁殖与呼吸综合征患猪样品,针对PRRSV的ORF5基因进行了遗传变异分析。通过实时荧光定量PCR鉴定PRRSV阳性样品和基因型,应用RT-PCR扩增PRRSV ORF5和ORF7基因,将阳性样本进行测序并开展系统发育学分群和GP5氨基酸突变分析。结果显示,共检出PRRSV-2阳性样品48份(总体阳性率约41%),经测序共获取38条ORF5序列和10条ORF7序列。所有ORF5序列间核苷酸相似性为77.1%~99.8%,ORF7序列间核苷酸相似性为83.3%~98.7%。系统发育学分群显示,谱系1占比60%(29/48,28株NADC30-like毒株,1株NADC34-like毒株),谱系3占比25%(12/48),谱系5.1占比5%(2/48),谱系8.7占比10%(5/48)。GP5氨基酸分析表明,信号肽(aa 1-26)、中和表位C(aa 52-61)及潜在的N-糖基化位点(aa 32-35、aa 44、aa 51)3个区域氨基酸存在显著变异。研究结果表明:多个谱系PRRSV同时在我国流行,谱系1已替代谱系8.7成为主要流行谱系,并分布于我国大多数省份;谱系3仅次于谱系1,主要在华南局部地区流行;谱系8.7毒株检出率较低。需要注意的是,在广东省惠州市检出1株NADC34-like毒株,说明该毒株可能已在华南地区潜在流行。近几年来,PRRSV在我国的流行日趋复杂,谱系1毒株的广泛流行增加了PRRSV的遗传多样性。本研究结果将为制定PRRSV防控策略提供参考。

关键词: 猪繁殖与呼吸综合征病毒, ORF5, 系统发育学分析, NADC30, NADC34

Abstract: In order to investigate the prevalence and genetic variation of porcine reproductive and respiratory syndrome virus (PRRSV) in China, our group collected 117 samples of clinically suspected PRRS symptoms from nearly 50 farms in 13 provinces in China from 2021 to 2022, and conducted genetic variation analysis for the ORF5 gene of PRRSV. The PRRSV-positive samples and genotypes were identified by Quantitative Real-time PCR, RT-PCR was applied to amplify PRRSV ORF5 and ORF7 genes, and the positive samples were sequenced and analyzed for phylogenetic analysis and GP5 amino acid mutations. The results showed that a total of 48 PRRSV-2 positive samples were detected (overall positive rate of about 41%). In addition, a total of 38 ORF5 sequences and 10 ORF7 sequences were obtained by sequencing. The nucleotide similarity among all ORF5 sequences ranged from 77.1% to 99.8%, and the nucleotide similarity among ORF7 sequences ranged from 83.3% to 98.7%. Phylogenetic analysis showed that lineage 1 accounted for 60% (29/48, 28 NADC30-like strains and 1 NADC34-like strain), lineage 3 for 25% (12/48), lineage 5.1 for 5% (2/48), and lineage 8.7 for 10% (5/48). Amino acid site analysis of GP5 showed significant amino acid variation in the signal peptide (aa 1-26), neutralizing epitope C (aa 52-61) and potential N-glycosylation sites (aa 32-35, aa 44, aa 51) were three regions with significant amino acid variants. The results of the study showed that several strains of PRRSV were prevalent in China at the same time, and lineage 1 had replaced lineage 8.7 as the main prevalent lineage and was distributed in most provinces in China; Lineage 3 was second only to lineage 1 and was prevalent mainly in the local area of southern China; Lineage 8.7 were detected at a low rate. It is important to note that a NADC34-like strain was detected in Huizhou City, Guangdong Province, indicating that this strain may have become potentially endemic in South China. The epidemic of PRRSV in China has become increasingly complex in recent years, and the widespread prevalence of lineage 1 strains has increased the genetic diversity of PRRSV. The results of this study will provide a reference for the development of PRRSV prevention and control strategies.

Key words: porcine reproductive and respiratory syndrome virus, ORF5, phylogenetic analysis, NADC30, NADC34

中图分类号:

S852.659.6

王志远, 刘博奇, 许志颖, 徐思佳, 邢家宝, 张桂红, 王衡, 孙彦阔. 2021—2022年我国部分地区猪繁殖与呼吸综合征病毒ORF5基因变异分析[J]. 畜牧兽医学报, 2023, 54(9): 3812-3823.

WANG Zhiyuan, LIU Boqi, XU Zhiying, XU Sijia, XING Jiabao, ZHANG Guihong, WANG Heng, SUN Yankuo. Analysis of ORF5 Gene Variation of Porcine Reproductive and Respiratory Syndrome Virus in Local Regions of China from 2021 to 2022[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(9): 3812-3823.

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2021—2022年我国部分地区猪繁殖与呼吸综合征病毒ORF5基因变异分析

Analysis of ORF5 Gene Variation of Porcine Reproductive and Respiratory Syndrome Virus in Local Regions of China from 2021 to 2022

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