一株PRRSV-2谱系1.8与1.5重组毒株的基因组特征分析

doi:10.11843/j.issn.0366-6964.2024.08.028

畜牧兽医学报 ›› 2024, Vol. 55 ›› Issue (8): 3570-3578.doi: 10.11843/j.issn.0366-6964.2024.08.028

一株PRRSV-2谱系1.8与1.5重组毒株的基因组特征分析

杨程¹(), 刘野¹, 程宁¹, 王凯月¹, 李欣蕾¹, 孙久英¹, 韩俊平², 李文军³, 王欢欢³, 邵笑³, 程雪娇⁴, 孙英峰¹^,*()

1. 天津农学院动物科学与动物医学学院, 天津 300384
2. 天津津垦牧业集团有限公司, 天津 300392
3. 天津农垦康嘉生态养殖有限公司, 天津 300386
4. 天津市中升挑战生物科技有限公司, 天津 300386

收稿日期:2023-09-20 出版日期:2024-08-23 发布日期:2024-08-28
通讯作者: 孙英峰 E-mail:164377125@qq.com;yfsun2000@163.com
作者简介:杨程(1998-)，女，广西梧州人，硕士生，主要从事动物传染病学研究，E-mail: 164377125@qq.com
基金资助:
天津市科技计划项目(22YFZCSN00100);天津市科技计划项目(22ZYCGSN00570);天津市科技计划项目(22YDTPJC00420)

Genomic Characterization of a Recombinant Strain of PRRSV-2 between Lineages 1.8 and 1.5

Cheng YANG¹(), Ye LIU¹, Ning CHENG¹, Kaiyue WANG¹, Xinlei LI¹, Jiuying SUN¹, Junping HAN², Wenjun LI³, Huanhuan WANG³, Xiao SHAO³, Xuejiao CHENG⁴, Yingfeng SUN¹^,*()

1. College of Animal Science and Animal Medicine, Tianjin Agricultural University, Tianjin 300384, China
2. Tianjin Jinken Animal Husbandry Group Co., Ltd, Tianjin 300392, China
3. Tianjin Nongken Kangjia Ecological Breeding Co., Ltd, Tianjin 300386, China
4. Tianjin Zhongsheng Challenge Biotechnology Co., Ltd, Tianjin 300386, China

Received:2023-09-20 Online:2024-08-23 Published:2024-08-28
Contact: Yingfeng SUN E-mail:164377125@qq.com;yfsun2000@163.com

摘要/Abstract

摘要：

为探究天津地区猪繁殖与呼吸综合征病毒(PRRSV)毒株的流行变异情况。本研究利用猪肺泡巨噬细胞(PAMs)分离培养、有限稀释法纯化和IFA鉴定，从具有严重呼吸症状断奶仔猪群中分离鉴定出一株新的PRRSV流行毒株(命名为TJ-C6)，并对其开展了全基因组扩增及其分子特征分析。结果显示，该毒株全基因组全长15011 bp，与NADC30毒株的核苷酸相似性最高(91.7%)，为NADC30-like PRRSV(谱系1.8)，且其Nsp 2存在131个氨基酸不连续缺失分子特征(“111+1+19 aa”)；GP5 N-糖基化位点分析显示存在3个糖基化位点(N32、N43、N50)。重组分析结果显示，该毒株是以NADC30-like(谱系1.8)为骨架，与NADC34-like(谱系1.5)发生重组的PRRSV，其重组区域位于12213~14628 nt(ORF2a~ORF6)。本研究分离鉴定到一株谱系1.8与谱系1.5重组的PRRSV-2毒株，可为天津地区的猪繁殖与呼吸综合征综合防控提供参考。

关键词: 猪繁殖与呼吸综合征病毒, 分离鉴定, 基因组特征, 重组

Abstract:

In order to investigate the epidemic variation of porcine reproductive and respiratory syndrome virus (PRRSV) strain in Tianjin area, a new epidemic strain of PRRSV (named TJ-C6) was isolated and identified from weaned piglets with severe respiratory symptoms by means of isolation and culture of porcine alveolar macrophages (PAMs), purification by limited dilution method and identification by IFA. The whole genome amplification and its molecular characteristics were analyzed. The results showed that the whole genome length of this strain was 15011bp, and the nucleotide homology with NADC30 strain was the highest (91.7%), which was NADC30-like PRRSV (lineage 1.8). Nsp2 had 131 discontinuous deletion features ("111+1+19aa"). GP5 N-glycosylation site analysis showed that there were three glycosylation sites (N32, N43, N50). Recombination analysis showed that this strain was a recombinant PRRSV with NADC30-like (lineage 1.8) as the skeleton and NADC34-like (lineage 1.5), and its recombination region was located in 12213-14628nt (ORF2a-ORF6). In this study, a recombinant PRRSV-2 strain of lineage 1.8 and lineage 1.5 was isolated and identified, which can provide reference for the prevention and control of porcine reproductive and respiratory syndrome in Tianjin.

Key words: PRRSV, isolation and identification, genomic characterization, recombination

中图分类号:

S852.659.6

杨程, 刘野, 程宁, 王凯月, 李欣蕾, 孙久英, 韩俊平, 李文军, 王欢欢, 邵笑, 程雪娇, 孙英峰. 一株PRRSV-2谱系1.8与1.5重组毒株的基因组特征分析[J]. 畜牧兽医学报, 2024, 55(8): 3570-3578.

Cheng YANG, Ye LIU, Ning CHENG, Kaiyue WANG, Xinlei LI, Jiuying SUN, Junping HAN, Wenjun LI, Huanhuan WANG, Xiao SHAO, Xuejiao CHENG, Yingfeng SUN. Genomic Characterization of a Recombinant Strain of PRRSV-2 between Lineages 1.8 and 1.5[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(8): 3570-3578.

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片段 Fragment	引物序列(5′→3′) Primer sequence	扩增片段大小/bp Amplified fragment size
1	F：ATCTATGCCACGACATTTGTATTG	1 568
	R：TCTGGAGACCTCACTCTTTCGG
2	F：GGCAGTGACTGACACGAGTGGAC	1 605
	R：TTTGTCGGGACACAGTTTTGC
3	F：GCCGTCGCCGATAGTGTCAA	1 498
	R：AGCACTTTTTACACCTACCTTG
4	F：CTGGACCACCCGTGTCTACT	1 621
	R：GGTGCTACACTGGGGGACA
5	F：CCAGTTGAATGGGTTGAAATC	1 225
	R：CCAGTTGAATGGGTTGAAAATC
6	F：TAGGACCCAAAAACCTTCGC	1 216
	R：GGCAAGGACTCTGGTCTCAA
7	F：GCCTGTCGGCAGCATCTTC	1 583
	R：GACGCTCTGAATGTCCTTGGT
8	F：CTGGAATAAACGGGAATAGG	1 558
	R：CCCACGACGAGTCTGATA
9	F：CGGGTCCTGTGATGAGTGTAA	1 527
	R：GAGCCTCGCCCCTGATAA
10	F：GCAGGTATCAGACTCGTCGTGG	1 522
	R：AAACCCCGACCACTGCGAC
11	F：AAGGAATCGGTCGCAGTGGT	1 576
	R：CCAGCGGAAACCAAAAACAG
12	F：TGAAGCCGAGACCTGTAAATA	1 529
	R：TTTTATTTAGCCAATCTGTGCC
13	F：CTTCAGTTATACAGCCCAGTTTC	1 485
	R：GTCATATACCCAAAAGTAAAGGC
14	F：CAGTTGATTTATAACCTGACGATA	1 549
	R：ATGGTTCTCGCCAATTAAATATTA

毒株 Strain	国家 Country	登录号 Accession No.	毒株 Strain	国家 Country	登录号 Accession No.
HLHDZD32-1901	China	MN648449	QYYZ	China	JQ308798.1
HLJZD30-1902	China	MN648055	GM2	China	JN662424.1
NADC34	USA	MF326985	CH-1R	China	EU807840
JL580	China	KR706343	CH-1a	China	AY032626
HENAN-XINX	China	KF611905	JXA1	China	EF112445
NADC30	USA	JN654459	JXA1-R	China	MT163314.1
CHsx1401	China	KP861625	TJ	China	EU860248
VR2332	USA	U87392	JXwn06	China	EF641008
RespPRRS MLV	USA	AF066183	ISU-10	USA	KT257966

病毒 Virus	断点 Breakpoints		主要亲本株 Major Parent	重组片段亲本株 Minor Parent	P值 P-value
病毒 Virus	开始 Beginning	结束 Ending	主要亲本株 Major Parent	重组片段亲本株 Minor Parent	RDP	GENECONV	Bootscan	MaxChi	Chimera	SiScan	Phylpro
TJ-C6	12 213	14 628	NADC34	NADC30	4.592× 10^-69	1.762×10^-43	5.230× 10^-65	1.080× 10^-20	6.431× 10^-17	3.720× 10^-20	1.811× 10^-13

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一株PRRSV-2谱系1.8与1.5重组毒株的基因组特征分析

Genomic Characterization of a Recombinant Strain of PRRSV-2 between Lineages 1.8 and 1.5

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