一株欧洲型猪繁殖与呼吸综合征病毒流行毒株的分子特征与致病性

doi:10.11843/j.issn.0366-6964.2025.12.030

摘要/Abstract

摘要：

猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndrome virus, PRRSV)分为欧洲型和美洲型两个种，即PRRSV-1和PRRSV-2，病毒存在基因变异，严重危害世界养猪业健康发展。本研究采用猪肺泡巨噬细胞(porcine alveolar macrophages, PAMs)从贵州省某发病猪场发病仔猪的肺脏组织分离获得1株PRRSV-1 GZ0308。该病毒株基因组全长14 970 bp，与PRRSV-1毒株LV相似性为86.99%，与PRRSV-2 VR2332相似性为66.49%。基因进化树分析，GZ0308毒株位于PRRSV-1的亚型1的一个新的分支。与LV毒株基因比较，GZ0308 NSP2基因编码区含有43个不连续氨基酸缺失，ORF3和ORF4的重叠区域存在5个连续氨基酸的缺失。5周龄仔猪以10^5.5TCID₅₀·mL^-1的病毒滴度分别肌肉和滴鼻接种GZ0308毒株病毒液各2 mL，结果显示，与对照组相比，GZ0308攻毒组仔猪出现短时体温升高、精神萎靡、日增重显著减少、肺脏出现典型弥漫性间质性肺炎病变，证明该毒株对仔猪具有致病性，从而为我国PRRSV分子流行病学研究提供了新的数据。

关键词: 猪繁殖与呼吸综合征病毒, 欧洲型, 分子特征, 致病性

Abstract:

Porcine reproductive and respiratory syndrome virus (PRRSV), comprising two distinct species, PRRSV-1 (European genotype) and PRRSV-2 (North American genotype), exhibits significant genetic variability and poses a substantial threat to global swine production. In this study, a PRRSV-1 strain, designated GZ0308, was isolated from the lung tissue of infected piglets on a farm in Guizhou Province using porcine alveolar macrophages (PAMs). The complete genome of GZ0308 spans 14 970 nucleotides, displaying 86.99% nucleotide identity with the PRRSV-1 reference strain LV and 66.49% with the PRRSV-2 strain VR2332. Phylogenetic analysis positioned GZ0308 within a novel branch of PRRSV-1 subtype 1. Comparative genomic analysis revealed that GZ0308 harbors 43 discontinuous amino acid deletions in the NSP2 coding region and a 5-amino-acid deletion in the overlapping region of ORF3 and ORF4 relative to the LV strain. To assess pathogenicity, five-week-old piglets were inoculated intramuscularly and intranasally with 2 mL of viral suspension (10^5.5TCID₅₀·mL^-1 per route). Compared to the control group, GZ0308-infected piglets exhibited transient pyrexia, lethargy, and a significant reduction in average daily weight gain. Postmortem examination identified characteristic diffuse interstitial pneumonia lesions, confirming the strain's pathogenicity in piglets. These findings contribute novel data to the molecular epidemiology of PRRSV in China and underscore the need for continued surveillance of emerging PRRSV variants.

Key words: PRRSV, the European type, molecular characteristics, pathogenicity

中图分类号:

S852.659.6

苏潼键, 张路捷, 孙杨杨, 张杰, 孙海凤, 姜平, 白娟. 一株欧洲型猪繁殖与呼吸综合征病毒流行毒株的分子特征与致病性[J]. 畜牧兽医学报, 2025, 56(12): 6281-6292.

SU Tongjian, ZHANG Lujie, SUN Yangyang, ZHANG Jie, SUN Haifeng, JIANG Ping, BAI Juan. Molecular Characteristics and Pathogenicity of an Epidemic Strain of European Type Porcine Reproductive and Respiratory Syndrome Virus[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(12): 6281-6292.

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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: ATGATGTGTAGGGTATTCCC	1 375
	R: CCATACCACTTATGCGCTC
2	F: ACCTGACTCTGGATGATGAT	2 005
	R: ATCATGTCAGGGAGGAATTTG
3	F: AGTCGTGCAACCCCAG	1 742
	R: AAAATAAAAATCCCCACCTCTCT
4	F: TGGTGTTCAAATCCTTTTTCATATC	1 979
	R: GGGGTGGCTGTTTCAG
5	F: ATGCTGCCGGCCAG	2 040
	R: GGGGTGCTGCGGTC
6	F: TCATGAAGAAGGCTTGGAAGT	1 960
	R: TGAGGGTCATAAATGAACAACCC
7	F: ATAACCATAGATTCATCCCAGGG	1 929
	R: GCCAGGTGTTGGAAATGAG
8	F: ATTGATGAGATGGTCTCTCGT	1 863
	R: TCCCCCGGTCGTC
9	F: CGGTACGTACTCTGCA	1 300
	R: TTTAATTTCGGTCACA

毒株 Strain	GenBank登录号 GenBank accesssion No.	基因型 Gene type	来源 Origin
Lelystad viurs	M96262	Subtype 1 PRRSV-1	荷兰Netherlands
Amervac	GU067771	Subtype 1 PRRSV-1	西班牙Spain
Cresa3266	JF276434	Subtype 1 PRRSV-1	西班牙Spain
DV-MLV	MT311646	Subtype 1 PRRSV-1	荷兰Netherlands
HKEU16	EU076704	Subtype 1 PRRSV-1	中国香港Hong Kong, China
FJQEU14	KP860913	Subtype 1 PRRSV-1	中国China
BJEU06-1	GU047344	Subtype 1 PRRSV-1	中国China
NMEU09-1	GU047345	Subtype 1 PRRSV-1	中国China
WestSib13	KX668221	Subtype 2 PRRSV-1	俄罗斯Russia
Lena	JF802085	Subtype 3 PRRSV-1	比利时Belgium
SU1-bel	KP889243	Subtype 3 PRRSV-1	白俄罗斯Belaurs
VR-2332	EF536003	PRRSV-2	美国USA

开放阅读框 Open reading frames	编码蛋白 Encode protein	位置 Region	碱基/bp Basic	氨基酸/aa Amino acid
ORF1a	NSP1	222-1 376	1 155	385
	NSP2	1 377-4 481	3 105	1 035
	NSP3	4 482-5 171	690	230
	NSP4	5 172-5 780	609	203
	NSP5	5 781-6 290	510	170
	NSP6	6 291-6 338	48	16
	NSP7α	6 339-6 785	447	149
	NSP7β	6 786-7 145	360	120
	NSP8	7 146-7 280	135	45
ORF1b	NSP9	7 265-9 199	1 935	684
	NSP10	9 200-10 524	1 325	442
	NSP11	10 526-11 198	672	224
	NSP12	11 198-11 653	456	152
ORF2	GP2	11 667-12 416	750	250
ORF3	GP3	12 275-13 072	798	266
ORF4	GP4	12 817-13 356	540	180
ORF5	GP5	13 353-13 958	606	201
ORF6	M	13 946-14 467	522	174
ORF7	N	14 457-14 843	387	129

序列 Sequence	LV		Amervac		DV-MLV		BJEU06-1		Lena		WestSib13
序列 Sequence	nt	aa	nt	aa	nt	aa	nt	aa	nt	aa	nt	aa
Full-length	86.99		85.88		86.88		84.98		80.88		80.74
5′UTR	95.93		94.57		95.93		93.67		87.74		90.50
ORF1a	86.74	86.86	85.30	85.27	86.62	87.52	84.22	83.61	80.33	76.91	77.86	77.16
ORF1b	87.07	95.36	86.08	95.22	86.98	95.29	85.03	95.22	80.67	91.59	80.94	91.87
ORF2	88.71	91.20	87.73	89.60	88.17	90.00	88.29	91.20	86.12	91.02	81.81	83.00
ORF3	87.24	83.40	86.19	82.26	87.36	84.15	87.06	82.38	81.78	76.31	82.27	80.59
ORF4	84.71	84.78	83.94	85.87	84.17	84.78	82.96	85.00	79.68	79.46	79.95	79.57
ORF5	84.56	86.14	84.49	86.63	84.32	86.14	82.21	84.65	82.72	80.69	78.22	77.23
ORF6	90.61	91.38	91.57	93.10	91.00	91.95	88.74	89.66	87.93	92.53	83.59	91.95
ORF7	90.05	90.70	88.11	91.47	90.05	90.70	89.66	87.60	86.45	87.10	83.29	83.06
3′UTR	95.28		92.98		95.28		94.74		93.86		91.89

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