Molecular Epidemiological Investigation and Genetic Evolution Analysis of ORF5 Gene of Porcine Reproductive and Respiratory Syndrome Virus in Henan Province in 2020

doi:10.15933/j.cnki.1004-3268.2022.03.017

Abstract

Abstract: In order to monitor the prevalence and variation of porcine reproductive and respiratory syndrome virus（PRRSV）in Henan Province，a total of 401 clinical samples of porcine reproductive and respiratory syndrome were collected in Henan Province in 2020 and analyzed by TaqMan real⁃time quantitative PCR（FQ⁃PCR）.The results showed that a total of 100 positive samples were detected，with a positive rate of 24.9%. The ORF5 genes of 18 PRRSV epidemic strains were sequenced for homologous alignment and genetic evolution analysis. Nucleotide homology analysis showed that the ORF5 gene nucleotide homology among 18 strains was above 83.4%，and the nucleotide homology with the reference strain was 82.9%—99.7%.The results of genetic evolution analysis showed that all the 18 PRRSV strains belonged to American genotype，including lineage 1，lineage 5 and lineage 8.Two PRRSV strains belonged to lineage 8 represented by JXAl，one PRRSV strain belonged to lineage 5 represented by VR2332 and 15 PRRSV strains belonged to lineage 1 represented by NADC30. The results of amino acid sequence analysis showed that the 13th amino acid virulence related site of HN⁃TH1⁃2020，HY⁃PY1⁃2020，HN⁃XY1⁃2020 and the 151st amino acid virulence related site of HN⁃XC1⁃2020 had different degrees of mutation.

Key words: Henan Province, Porcine reproductive and respiratory syndrome virus, Epidemiology, ORF5 gene, Variation

摘要： 为了解河南省猪繁殖与呼吸综合征病毒（PRRSV）的流行及变异情况，对2020年采集于河南的401份PRRSV感染疑似病料进行TaqMan荧光定量PCR（FQ-PCR）检测。结果显示，共检测出100份阳性样品，阳性率为24.9%。对其中18株PRRSV流行毒株ORF5基因进行测序、同源性比对和遗传进化分析。同源性比对结果显示，18株PRRSV流行毒株ORF5基因核苷酸序列同源性在83.4%以上，与参考毒株同源性为82.9%~99.7%。遗传进化分析结果显示，18株PRRSV流行毒株序列均为美洲株，分属于1、5、8三个谱系，其中2株属于以JXAl为代表的谱系8，1株属于以VR2332为代表的谱系5，15株属于以NADC30为代表的谱系1。氨基酸比对结果显示，HN-TH1-2020、HY-PY1-2020、HN-XY1-2020毒株的13位以及HN-XC1-2020毒株的151位氨基酸毒力相关位点发生了不同程度的突变。

关键词: 河南省, 猪繁殖与呼吸综合征病毒, 分子流行病学, ORF5基因, 变异

CLC Number:

S855.3

GUO Zhanda, DU Jimei, GUO Ziyi, WANG Xingang, ZHANG Yanhua. Molecular Epidemiological Investigation and Genetic Evolution Analysis of ORF5 Gene of Porcine Reproductive and Respiratory Syndrome Virus in Henan Province in 2020[J]. Journal of Henan Agricultural Sciences, 2022, 51(3): 146-153.

郭占达, 杜季梅, 郭子仪, 王新港, 张彦华. 2020 年河南省猪繁殖与呼吸综合征病毒的分子流行病学调查及ORF5 基因变异分析[J]. 河南农业科学, 2022, 51(3): 146-153.

References

［1］陈溥言.兽医传染病学［M］.6版. 北京：中国农业大学出版社，2015：223⁃228.
CHEN P Y.Veterinary lemology［M］.6th edition. Beijing：China Agricultural University Press，2015：223⁃228.
［2］樊振华，刘文俊，武守艳，等.猪繁殖与呼吸综合征病毒山西分离株ORF7基因序列分析［J］.山西农业科学，2020，48（3）：446⁃448.
FAN Z H，LIU W J，WU S Y，et al.Sequence analysis on the ORF7 gene of Shanxi isolate porcine reproductive and
respiratory syndrome virus ［J］.Journal of Shanxi Agricultural Sciences，2020，48（3）：446⁃448.
［3］WENSVOORT G，TERPSTRA C，POL J M A，et al.Mystery swine disease in the Netherlands：The isolation of Lelytad virus［J］.Vet Dermatol，1991，13（3）：121⁃130.
［4］SNIJDER E J，KIKKERT M，FANG Y.Arterivirus molecular biology and pathogenesis［J］.J Genvirol，2013，94（10）：2141⁃2163.
［5］郭宝清，陈章水，刘文兴，等.从疑似PRRS流产胎儿分离PRRSV的研究［J］.中国畜禽传染病学，1996（2）：1⁃5.
GUO B Q，CHEN Z S，LIU W X，et al.Isolation and Identification of porcine reproductive and respiratory syndrome（PRRS）virus from aborted fetuses suspected of PRRS［J］.Chinese Journal of Infectious Disease，1996（2）：1⁃5.
［6］FENG Y， ZHAO T， NGUYEM T，et al.Porcine respiratory and reproductive syndrome virus variants，Vietnam and China，2007［J］.Emerg Infect Dis，2008，14（11）：1774⁃1176.
［7］FORSBERG R. Divergence time of porcine reproductive and respiratory syndrome virus subtypes［J］.Molecular Biology and Evolution，2005，22（11）：2131⁃2134.
［8］SHI M，LAM T T，HON C C，et al.Molecular epidemiology of PRRSV：A phylogenetic perspective［J］.Virus Research，2010，154（1/2）：7⁃17.
［9］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⁃15，18.
QU X Y，ZHANG 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⁃15，18.
［10］周峰.2012—2013年河南地区猪繁殖与呼吸综合征病毒分子流行病学调查及河南流行株的分离、鉴定［D］.郑州：河南农业大学，2014.
ZHOU F.Molecular epidemiology survey，isolation and identification of the porcine reproductive and respiratory
syndrome virus in Henan province from 2012 to 2013［D］.Zhengzhou：Henan Agricultural University，2014.
［11］ZHAO K，YE C，CHANG X B，et al.Importation and recombination are responsible for the latest emergence
of highly pathogenic porcine reproductive and respiratory syndrome virus in China［J］.Journal of Virology，2015，89（20）：107⁃116.
［12］OSTROWSKI M，GALEOTA J A，JAR A M，et al.Identification of neutralizing and nonneutralizing epitopes in the porcine reproductive and respiratory syndrome virus GP5 ectodomain［J］.Journal of Virology，2002，76（9）：4241⁃4250.
［13］PLAGEMANN P G.The primary GP5 neutralization epitope of North American isolates of porcine reproductive and respiratory syndrome virus ［J］.Veterinary Immunology and Immunopathology，2004，102（3）：263⁃275.
［14］GAO Z Q， GUO X， YANG H C.Genomic characterization of two Chinese isolates of porcine respiratory and reproductive syndrome virus［J］.Arch Virol，2004，149（7）：41⁃51.
［15］TIAN K，YU X，ZHAO T，et al.Emergence of fatal PRRSV variants：Unparalleled outbreaks of atypical PRRS in China and molecular dissection of the unique allmark［J］.PLoS One，2007，2（6）：526.
［16］ZHOU L，WANG Z，DING Y，et al.NADC30⁃like strain of porcine respiratory and reproductive syndrome virus，China［J］.Emerg Infect Dis，2015，21（12）：2256⁃2257.
［17］杨汉春，周磊. 猪繁殖与呼吸综合征病毒的遗传变异与演化［J］.生命科学，2016，28（3）：325⁃336.
YANG H C，ZHOU L.Genetic variation and evolution of porcine reproductive and respiratory syndrome virus［J］.Chinese Bulletin of Life Sciences，2016，28（3）：325⁃336.
［18］郭天准.2014—2015年河南省PRRSV的分子检测、变异分析及HP-PRRSV分离株全基因组序列分析［D］.郑州：河南农业大学，2016.
GUO T Z.Molecular identification and variation analysis of PRRSV in Henan province from 2014 to 2015 and analysis of complete genome sequences of HP⁃PRRSV isolates［D］.Zhengzhou：Henan Agricultural University，2016.
［19］王新港.猪繁殖与呼吸综合征病毒分离鉴定、变异与重组及致病性研究［D］.郑州：河南农业大学，2018.
WAGN X G.Isolation， identification，variation，recombination and pathogenicity of porcine reproductive and respiratory syndrome viruses［D］.Zhengzhou：Henan Agricultural University，2018.
［20］赵小月.2018—2019年河南地区猪繁殖与呼吸综合征病毒分子流行病学调查［D］.郑州：河南农业大学，2020.
ZHAO X Y.Molecular epidemiology of the porcine reproductive and respiratory syndrome virus in Henan province from 2018 to 2019［D］.Zhengzhou：Henan Agricultural Univercity，2020.
［21］陈盛楠，张海龙，谢博，等.2018年广东部分地区猪繁殖与呼吸综合征病毒流行毒株ORF5基因变异分析［J］.动物医学进展，2020，41（1）：50⁃56.
CHEN S N，ZHANG H L，XIE B，et al.Genetic variation analysis of ORF5 gene of porcine reproductive and respiratory syndrome virus in Guangdong Province in 2018［J］.Progress in Veterinary Medicine，2020，41（1）：50⁃56.
［22］王雅婷，柴娟，张红丽，等.浙江省猪繁殖与呼吸综合征病毒ORF5基因的遗传变异分析［J］.中国畜牧兽医，2019，46（12）：3715⁃3724.
WANG Y T，CHAI J，ZHANG H L，et al.Genetic variation analysis of ORF5 gene of porcine reproductive and respiratory syndrome virus in Zhejiang Province［J］.Veterinary Medicine，2019，46（12）：3715⁃3724.

[1]	FANG Jianyu, ZHANG Qingxian, LANG Limin, XU Bin, WANG Gaili, XI Yanyan, FENG Xianming, WANG Keling, LI Shaoyu. Prokaryotic Expression and Anti‑PRRSV Activity Study of Porcine IFN‑α8 [J]. Journal of Henan Agricultural Sciences, 2023, 52(9): 141-147.
[2]	HU Feng, ZHANG Jiantao, ZHANG Jie, GAO Tongmei, ZHAO Qiaoli, ZHENG Guoqing, LI Guoqiang, LIU Lijie. Study on Spatio‑Temporal Variation of Climatic Suitability of Sesame Planting in Henan Province [J]. Journal of Henan Agricultural Sciences, 2023, 52(8): 56-68.
[3]	XIN Yujie, LIU Jihong, FENG Dan, ZHAO Guanghua, PEI Jinhua, YIN Haiyan, WANG Yun, WANG Hongqi, LIU Dongmei, CAO Cheng, WANG Junyan, ZHENG Jia, CAO Yingni. Quality Character Analysis of Wheat Varieties（Lines）in Henan Regional Tests from 2017 to 2021 [J]. Journal of Henan Agricultural Sciences, 2023, 52(2): 31-37.
[4]	RONG Yasi, LI Guoqiang, ZHANG Jie, ZHANG Jiantao, WANG Meng, ZHENG Guoqing, FENG Wei. Irrigation Strategies Optimization for Winter Wheat in Henan Province Based on AquaCrop Model [J]. Journal of Henan Agricultural Sciences, 2023, 52(2): 151-161.
[5]	LI Jinhua, SHU Wentao, CHANG Shihao, YANG Qingchun, ZHANG Donghui, ZHANG Baoliang, GENG Zhen. Analysis of Yield and Quality of Soybean Varieties in Regional Trial in Henan Province from 1985 to 2020 [J]. Journal of Henan Agricultural Sciences, 2022, 51(1): 50-59.
[6]	BAI Jingke, LIU Fang, SONG Ruifang, LI Chengjun, CHEN Yuguo, WANG Xuefen, ZHANG Zhaoyang, WANG Zichen, CUI Guojie, LI Shujun. Population Dynamics of Winged Aphids in Tobacco Planting Areas of Henan Province [J]. Journal of Henan Agricultural Sciences, 2021, 50(5): 80-85.
[7]	PENG Zhifeng, HUANG Hui, ZHANG Xiaozhan, JIANG Zenghai, SHI Hongtao, QIAO Hongxing, BIAN Chuanzhou. Identification and Susceptibility of Actinobacillus pleuropneumoniae from Henan Province [J]. Journal of Henan Agricultural Sciences, 2021, 50(1): 158-163.
[8]	ZHANG Jie, ZHANG Hui, CHEN Dandan, LI Guoling, ZHANG Jiantao, ZANG Hecang, ZHAO Qing, HU Feng, LI Guoqiang, ZHENG Guoqing. Spatial and Temporal Distribution Characteristics of Effective Rainfall and Water Requirement of Summer Maize Growth Period in Henan Province [J]. Journal of Henan Agricultural Sciences, 2020, 49(9): 173-180.
[9]	FENG Lili, DUAN Junzhi, YANG Cuiping, LIU Yuxia, SUN Yan, ZHANG Huifang, YAN Zhaoling, CHEN Haiyan, QI Hongzhi, ZHUO Wenfei. Application of Agricultural Big Data in Pig Production in Henan Province [J]. Journal of Henan Agricultural Sciences, 2020, 49(7): 155-160.
[10]	WANG Shangkun, HUANG Qian, LI Zhengli, WANG Zilin. Comparative Test on the Scale Cultivation of Different Varieties of Red Pear [J]. Journal of Henan Agricultural Sciences, 2020, 49(5): 118-125.
[11]	LI Jiaxiao, WANG Xianwei, LI Zhiming, ZHANG Zijing, LIU Xian, HE Hua, WANG Eryao, XU Zejun, RU Baorui, CHEN Hong, HUANG Yongzhen. Thoughts on Selection and Breeding and Protective Countermeasures of Genetic Resources of Five Native Goat Breeds in Henan Province [J]. Journal of Henan Agricultural Sciences, 2020, 49(5): 168-172.
[12]	HE Shuyan, ZHANG Long. A New Record Genus and Species of Lampyridae from Henan Province [J]. Journal of Henan Agricultural Sciences, 2020, 49(12): 105-106.
[13]	LI Shimin, ZHANG Hongli, WANG Laigang, ZHENG Guoqing, GUO Yan, GAO Jianhua. Winter Wheat Agrometeorological Drought Zoning and Analysis in Henan Province Based on Grid and Fuzzy Clustering Algorithm [J]. Journal of Henan Agricultural Sciences, 2020, 49(11): 172-180.
[14]	QIN Feng, LIU Yun, WU Zhi, WU Shuang, GUO Changming, GU Lingling, WU Xiaojie, ZHU Shanyuan. Anti-porcine Reproductive and Respiratory Syndrome Virus Effect of Extracts of Valeriana jatamansi Jones [J]. Journal of Henan Agricultural Sciences, 2020, 49(10): 149-155.
[15]	SHEN Yunting, SUO Yanyan, ZHANG Xiang, SI Xianzong, YU Qiong, YU Hui, SUN Yanmin, WU Jihua, CHEN Lei. Evaluation of Soil Nutrient Status and Suggestions for Fertilization in Main Peanut Production Areas of Henan Province [J]. Journal of Henan Agricultural Sciences, 2019, 48(9): 67-73.