猪繁殖与呼吸综合征病毒湖南分离株ORF5基因的遗传进化分析

doi:10.16431/j.cnki.1671-7236.2021.10.032

Abstract

Abstract: In order to explore infection of Porcine reproductive and respiratory syndrome virus (PRRSV) in native pig breeding farms of Hunan province, 287 blood samples were collected from two native pig breeding farms during 2019 to 2020. PRRSV were detected by RT-PCR in 41 mixed seral samples. Open reading frame 5 (ORF5) gene of PRRSV were amplified from PRRSV positive samples by high fidelity PCR. DNAStar software was performed to analyze the genetic evolution of ORF5 gene and GP5 amino acid between these PRRSV strains and different PRRSV strains from China and abroad after sequencing. Finally, PRRSV positive seral were inoculated in Marc-145 cells followed by blind passaging. The titer of the isolated PRRSV was determined by Reed-Muench method. The results showed that 3 samples from the 41 mixed samples were positive for PRRSV. Six ORF5 sequences were amplified from PRRSV positive mixed samples, all of which belonged to branch lineage 8 of PRRSV-2 strain, and the nucleotide homology was 99.2% to 99.8%. The corresponding GP5 protein acid sequences encoded by six ORF5 gene sequences were different in the signal peptide region (position 23), potential N-glycosylation site (position 33) and epitope C (position 59). Obvious cytopathic effect (CPE) was observed in the Marc-145 cells inoculated the PRRSV positive sera after five blind passages. A stain of PRRSV designated as NX-1 was obtained with a TCID₅₀ of 4×10⁵/mL. The data indicated that there existed infection of PRRSV in Hunan native pig breeding farms. The prevalent PRRSV isolate belonged to lineage 8 of PRRSV-2 with several variations in the amino acid sequence of GP5, which might be associated with immune failure. The present study could supply a foundation for immune prevention and control in Hunan native pig breeding farms.

Key words: porcine reproductive and respiratory syndrome (PRRSV); ORF5 gene; genetic evolution analysis

CLC Number:

S852.65⁺1

DENG Jiping, CHEN Zhilong, GUO Songyangnian, ZHANG Kun, XU Daojun, YANG Qing, YUAN Anwen. Genetic Evolution Analysis of ORF5 Gene of Porcine Reproductive and Respiratory Syndrome Virus Isolated from Hunan Province[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(10): 3795-3802.

References

[1] ZHOU Y, CHEN L, ZHANG L, et al.Simultaneous identification of 6 pathogens causing porcine reproductive failure by using multiplex ligation-dependent probe amplification[J]. Transboundary and Emerging Diseases, 2020, 67(6):2467-2474.
[2] YIN B, QI S, SHA W, et al.Molecular characterization of the Nsp2 and ORF5(ORF5a) genes of PRRSV strains in nine provinces of China during 2016-2018[J]. Frontiers in Veterinary Science, 2021, 8:605832.
[3] KAPPES M A, FAABERG K S.PRRSV structure, replication and recombination:Origin of phenotype and genotype diversity[J]. Virology, 2015, 479-480:475-486.
[4] SHI M, LAM T T, HON C C, et al.Phylogeny-based evolutionary, demographical, and geographical dissection of North American type 2 Porcine reproductive and respiratory syndrome viruses[J]. Journal of Virology, 2010, 84(17):8700-8711.
[5] CHEN N, XIAO Y, YE M, et al.High genetic diversity of Chinese Porcine reproductive and respiratory syndrome viruses from 2016 to 2019[J]. Research in Veterinary Science, 2020, 131:38-42.
[6] POPESCU L N, TRIBLE B R, CHEN N, et al.GP5 of Porcine reproductive and respiratory syndrome virus (PRRSV) as a target for homologous and broadly neutralizing antibodies[J]. Veterinary Microbiology, 2017, 209:90-96.
[7] HOU J, LI R, QIAO S, et al.Glycoprotein 5 is cleaved by cathepsin E during Porcine reproductive and respiratory syndrome virus membrane fusion[J]. Journal of Virology, 2020, 94(10):e00097-20.
[8] FERNANDEZ A, SUAREZ P, CASTRO J M, et al.Characterization of regions in the GP5 protein of Porcine reproductive and respiratory syndrome virus required to induce apoptotic cell death[J]. Virus Research, 2002, 83(1-2):103-118.
[9] 潘玉春.加快推进地方猪遗传资源保护与利用[J]. 中国畜牧业, 2020, 6:25-27. PAN Y C.Accelerate the protection and utilization of local pig genetic resources[J]. China Animal Industry, 2020, 6:25-27.(in Chinese)
[10] 陈指龙, 邓吉平, 湛洋, 等.猪圆环病毒2型湖南株的分离鉴定与进化分析[J]. 中国兽医科学, 2020, 50(12):1522-1530. CHEN Z L, DENG J P, ZHAN Y, et al.Identification and genetic variation analyses of Porcine circovirus type 2 isolated from Hunan province[J]. Chinese Veterinary Science, 2020, 50(12):1522-1530.(in Chinese)
[11] 王婕敏.猪精液中PRRSV、CSFV、JEV荧光定量PCR检测方法的建立及初步应用[D].长沙:湖南农业大学, 2016. WANG J M.Establishment and application of fluorescent quantitative PCR methods for detection of PRRSV, CSFV and JEV in boar semen[D].Changsha:Hunan Agricultural University, 2016.(in Chinese)
[12] 冷雪.高致病性猪繁殖与呼吸综合征病毒分离鉴定、遗传变异及致病性分析[D].北京:中国农业科学院, 2012. LENG X.Isolation, identification, genetic mutation and pathogenicity analysis of highly Porcine reproductive and respiratory syndrome virus[D].Beijing:Chinese Academy of Agricultural Sciences, 2012.(in Chinese)
[13] 施开创, 尹彦文, 王孝德, 等.2018年广西重要猪源病毒性疫病流行病学调查[J]. 中国畜牧兽医, 2020, 47(1):174-181. SHI K C, YIN Y W, WANG X D, et al.Epidemiological investigation of major porcine viral diseases in Guangxi during 2018[J]. China Animal Husbandry & Veterinary Medicine, 2020, 47(1):174-181.(in Chinese)
[14] 孙秋艳, 孙霞, 王志远.2014-2016年山东省规模化养猪场猪常见病毒性疾病流行病学调查[J]. 黑龙江畜牧兽医, 2019, 2:72-75. SUN Q Y, SUN X, WANG Z Y.Epidemiological survey of common viral diseases in pigs on large-scale pig farms in Shandong province from 2014 to 2016[J]. Heilongjiang Animal Science and Veterinary Medicine, 2019, 2:72-75.(in Chinese)
[15] LIANG W, ZHAO T, PENG Z, et al.Epidemiological and genetic characteristics of Porcine reproductive and respiratory syndrome virus circulating in Central and South China in 2016[J]. Acta Tropica, 2019, 190:83-91.
[16] GUO Z, CHEN X X, LI R, et al.The prevalent status and genetic diversity of Porcine reproductive and respiratory syndrome virus in China:A molecular epidemiological perspective[J]. Virology Journal, 2018, 15(1):2.
[17] GUO Z, CHEN X, LI X, et al.Prevalence and genetic characteristics of Porcine reproductive and respiratory syndrome virus in Central China during 2016-2017:NADC30-like PRRSVs are predominant[J]. Microbial Pathogenesis, 2019, 135:103657.
[18] 朱国坡, 马凤龙, 王家福, 等.2016-2018年我国华北部分地区猪繁殖与呼吸综合征病毒ORF5基因的遗传变异分析[J]. 黑龙江畜牧兽医, 2020, 11:96-100. ZHU G P, MA F L, WANG J F, et al.Genetic evolution analysis of ORF5 gene of Porcine reproductive and respiratory syndrome virus in some areas of North China in the spring from 2016 to 2018[J]. Heilongjiang Animal Science and Veterinary Medicine, 2020, 11:96-100.(in Chinese)
[19] 曲向阳, 孙英军, 张洪亮, 等.2014-2017年山东省部分规模化猪场PRRSV ORF5基因的遗传演化分析[J]. 中国兽医杂志, 2018, 54(9):12-15. QU X Y, SUN Y J, ZHANG H L, et al.Genetic variation and phylogenetic analysis of PRRSV ORF5 gene from large scale pig farms in Shandong province during 2014-2017[J]. Chinese Journal of Veterinary Medicine, 2018, 54(9):12-15.(in Chinese)
[20] SUN Y K, LI Q, YU Z Q, et al.Emergence of novel recombination lineage 3 of Porcine reproductive and respiratory syndrome viruses in Southern China[J]. Transboundary and Emerging Diseases, 2018, 66(1):578-587.
[21] XIE S, LIANG W, WANG X, et al.Epidemiological and genetic characteristics of Porcine reproduction and respiratory syndrome virus 2 in mainland China, 2017-2018[J]. Archives of Virology, 2020, 165(7):1621-1632.
[22] 刘瀚扬, 张毅, 肖冉, 等.2018年四川地区猪繁殖与呼吸综合征病毒ORF5基因的分子流行病学调查研究[J]. 中国畜牧兽医, 2020, 47(4):1172-1182. LIU H Y, ZHANG Y, XIAO R, et al.Molecular epidemiological investigation on ORF5 gene of Porcine reproductive and respiratory syndrome virus in Sichuan in 2018[J]. China Animal Husbandry & Veterinary Medicine, 2020, 47(4):1172-1182.(in Chinese)
[23] MARDASSI H, MASSIE B, DEA S.Intracellular synthesis, processing, and transport of proteins encoded by ORFs 5 to 7 of Porcine reproductive and respiratory syndrome virus[J]. Virology, 1996, 221(1):98-112.
[24] THAA B, SINHADRI B C, TIELESCH C, et al.Signal peptide cleavage from GP5 of PRRSV:A minor fraction of molecules retains the decoy epitope, a presumed molecular cause for viral persistence[J]. PLoS One, 2013, 8(6):e65548.
[25] WEI Z, LIN T, SUN L, et al.N-linked glycosylation of GP5 of Porcine reproductive and respiratory syndrome virus is critically important for virus replication in vivo[J]. Journal of Virology, 2012, 86(18):9941-9951.
[26] WEI C, DAI A, FAN J, et al.Efficacy of type 2 PRRSV vaccine against challenge with the Chinese lineage 1(NADC30-like) PRRSVs in pigs[J]. Scientific Reports, 2019, 9:10781.
[27] BAI X, WANG Y, XU X, et al.Commercial vaccines provide limited protection to NADC30-like PRRSV infection[J]. Vaccine, 2016, 34(46):5540-5545.

Genetic Evolution Analysis of ORF5 Gene of Porcine Reproductive and Respiratory Syndrome Virus Isolated from Hunan Province

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