2018-2020年中国部分省市猪流行性腹泻病毒S1基因的监测及遗传变异分析

doi:10.16431/j.cnki.1671-7236.2022.02.003

摘要/Abstract

摘要： 【目的】调查中国规模化猪场猪流行性腹泻病毒(Porcine epidemic diarrhea virus，PEDV)的流行情况。【方法】利用RT-PCR检测方法对2018-2020年采集于全国11个省市318个规模化猪场的2 391份样品进行PEDV核酸检测，对30份阳性样品进行S1全基因扩增与测序，并利用MegAlign、Mega 7.0等软件进行分析。【结果】 2018-2020年，猪场PEDV核酸阳性率分别为48.57%、10.96%和4.72%，样品PEDV核酸阳性率分别为28.97%、8.27%和7.50%，猪场和样品PEDV核酸阳性率呈现逐年下降的趋势。S1基因相似性分析显示，获得的30株毒株全部为GⅡ基因型，其中17株毒株为GⅡa基因亚型、5株毒株为GⅡb亚型、8株毒株为GⅡc亚型，GⅡa基因亚型毒株为2018-2020年流行的主要毒株类型；30株毒株S1基因序列之间核苷酸相似性为93.0%~99.5%, 氨基酸相似性为91.7%~100%，其中22株毒株与2017-2018年流行的GⅡ基因型毒株相比S1氨基酸序列表现出特征性插入和缺失。【结论】本研究结果为监测和分析中国PEDV的变异和演化提供了临床数据，为猪流行性腹泻防控和疫苗研制提供了参考。

关键词: 猪流行性腹泻病毒(PEDV); S1基因; 遗传变异分析

Abstract: 【Objective】 This study was aimed to survey the prevalence of Porcine epidemic diarrhea virus (PEDV) in large-scale pig farms in China.【Method】 2 391 samples collected from 318 large-scale pig farms in 11 provinces and municipalities of China from 2018 to 2020 were tested for PEDV nucleic acid using RT-PCR.The S1 gene of 30 positive samples were amplified and sequenced, and analyzed by MegAlign, Mega 7.0 and other softwares.【Result】 From 2018 to 2020, the PEDV nucleic acid positive rates in pig farms were 48.57%, 10.96%, and 4.72%, respectively, and the PEDV nucleic acid positive rates in samples were 28.97%, 8.27%, and 7.50%, respectively.The positive rate of PEDV nucleic acid in pig farms and samples showed a downward trend year by year.The similarity analysis of S1 gene showed that all the 30 strains obtained in this experiment were GⅡ genotype, of which 17 strains were GⅡa subtype, 5 strains were GⅡb subtype, and 8 strains were GⅡc subtype, GⅡa subtype strains were the main types of strains circulating from 2018 to 2020.The nucleotide similarity among the 30 strains sequence was 93.0%-99.5%, and the amino acid similarity was 91.7%-100%, compared with the GⅡ genotype strains prevalent from 2017 to 2018, the S1 amino acid sequences of 22 strains showed characteristic insertion and deletion.【Conclusion】 The results provided clinical data support for surveying and analyzing the variation and evolution of PEDV in China, and provided reference for porcine epidemic diarrhea prevention and control and vaccine development.

Key words: Porcine epidemic diarrhea virus(PEDV); S1 gene; genetic variation analysis

中图分类号:

S852.65⁺1

颜忠, 杨汉春. 2018-2020年中国部分省市猪流行性腹泻病毒S1基因的监测及遗传变异分析[J]. 中国畜牧兽医, 2022, 49(2): 424-434.

YAN Zhong, YANG Hanchun. Surveillance and Genetic Variation Analysis of S1 Gene of Porcine Epidemic Diarrhea Virus in Part Provinces and Municipalities of China from 2018 to 2020[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(2): 424-434.

参考文献

[1] JUNG K, SAIF L J. Porcine epidemic diarrhea virus infection: Etiology, epidemiology, pathogenesis and immunoprophylaxis[J]. Veterinary Journal, 2015, 204(2): 134-143.
[2] WOOD E N. An apparently new syndrome of porcine epidemic diarrhoea[J]. The Veterinary Record, 1977, 100(12): 243-244.
[3] PENSAERT M B, DE BOUCK P. A new coronavirus-like particle associated with diarrhoea in swine[J]. Archives of Virology, 1978, 58(3): 243-247.
[4] CIMA G. Fighting a deadly pig disease. Industry, veterinarians trying to contain PED virus, new to the US[J]. Jounral of the American Veterinary Medical Association, 2013, 243(4): 467-470.
[5] PARK S J, SONG D S, HA G W, et al. Cloning and further sequence analysis of the spike gene of attenuated Porcine epidemic diarrhea virus DR13[J]. Virus Genes, 2007, 35(1): 55-64.
[6] SUN D B, FENG L, SHI H Y, et al. Spike protein region (aa 636-789) of Porcine epidemic diarrhea virus is essential for induction of neutralizing antibodies[J]. Acta Virologica, 2007, 51(3): 149-156.
[7] CHEN Q, LI G, STASKO J, et al. Isolation and characterization of Porcine epidemic diarrhea viruses associated with the 2013 disease outbreak among swine in the United States[J]. Jounral of Clinical Microbiology, 2014, 52: 234-243.
[8] GUO J H, FANG L R, YE X, et al. Evolutionary and genotypic analyses of global Porcine epidemic diarrhea virus strains[J]. Transboundary and Emerging Diseases, 2019, 66(1): 111-118.
[9] 熬超杰. 猪流行性腹泻病毒遗传变异与进化分析[D]. 武汉: 华中农业大学, 2019.
AO C J. Genetic variation and evolutionary analysis of Porcine epidemic diarrhea virus[D]. Wuhan: Huazhong Agricultural University, 2019. (in Chinese)
[10] 张志, 董雅琴, 刘爽, 等. 我国部分省份猪流行性腹泻的流行病学监测[J]. 中国动物检疫, 2014, 31(10): 47-51.
ZHANG Z, DONG Y Q, LIU S, et al. Survey and surveillance of porcine epidemic diarrhea in some provinces in China[J]. China Animal Health Inspection, 2014, 31(10): 47-51. (in Chinese)
[11] 蒋新华, 周荷田, 罗锋, 等. 2017年江西部分地区PEDV分子流行病学调查[J]. 中国畜牧兽医, 2019, 46(5): 1474-1482.
JIANG X H, ZHOU H T, LUO F, et al. Molecular epidemiological investigation of Porcine epidemic diarrhea virus in Jiangxi province in 2017[J]. China Animal Husbandry & Veterinary Medicine, 2019, 46(5): 1474-1482. (in Chinese)
[12] 严瑾, 施开创, 刘宏梅, 等. 2017-2019年广西猪重要病毒性腹泻病原流行病学调查[J]. 广西农学报, 2020, 35(4): 20-25.
YAN J, SHI K C, LIU H M, et al. Epidemiological investigation of major porcine viral diarrhea pathogens in Guangxi from 2017 to 2019[J]. Journal of Guangxi Agriculture, 2020, 35(4): 20-25. (in Chinese)
[13] 袁翠霞, 景小金, 陈红, 等. 2018-2019年贵阳市部分规模化猪场主要病毒性腹泻的流行学调查[J]. 养猪, 2020, 4: 119-120.
YUAN C X, JING X J, CHEN H, et al. Epidemiological survey of major viral diarrhea in part large-scale pig farms in Guiyang from 2018 to 2019[J]. Swine Production, 2020, 4: 119-120. (in Chinese)
[14] TAN L, LI Y, HE J, et al. Epidemic and genetic characterization of Porcine epidemic diarrhea virus strains circulating in the regions around Hunan, China, during 2017-2018[J]. Archives of Virology, 2020, 165: 877-889.
[15] WANG Q, VLASOVA A N, KENNEY S P, et al. Emerging and re-emerging Coronaviruses in pigs[J]. Current Opinion in Virology, 2019, 34: 39-49.
[16] 谢荣辉, 赵灵燕, 倪柏峰, 等. 浙江省猪流行性腹泻病毒S1基因克隆与序列分析[J]. 中国畜牧兽医, 2018, 45(6): 1668-1676.
XIE R H, ZHAO L Y, NI B F, et al. Cloning and sequence analysis of S1 gene of Porcine epidemic diarrhea virus from Zhejiang province[J]. China Animal Husbandry & Veterinary Medicine, 2018, 45(6): 1668-1676. (in Chinese)
[17] LI W, LI H, LIU Y, et al. New variants of Porcine epidemic diarrhea virus, China, 2011[J]. Emerging Infectious Diseases, 2012, 18(8): 1350-1353.
[18] SUN R Q, CAI R J, CHEN Y Q, et al. Outbreak of porcine epidemic diarrhea in suckling piglets, China[J]. Emerging Infectious Diseases, 2012, 18(1): 161-163.
[19] WANG X M, NIU B B, YAN H, et al. Genetic properties of endemic Chinese Porcine epidemic diarrhea virus strains isolated since 2010[J]. Archives of Virology, 2013, 158(12): 2487-2494.
[20] AYUDHYAY S N, ASSAVACHEEP P, THANA-WONGN-UWECH R. One world-one health: The threat of emerging swine diseases. An Asian perspective[J]. Transboundary and Emerging Diseases, 2012, 59(1): 9-17.
[21] LEE S, LEE C. Outbreak-related Porcine epidemic diarrhea virus strains similar to US strains, South Korea, 2013[J]. Emerging Infectious Diseases, 2014, 20(7): 1223-1226.
[22] LEE D K, PARK C K, KIM S H, et al. Heterogeneity in spike protein genes of Porcine epidemic diarrhea viruses isolated in Korea[J]. Virus Research, 2010, 149(2): 175-182.
[23] KIM S H, LEE J M, JUNG J, et al. Genetic characterization of Porcine epidemic diarrhea virus in Korea from 1998 to 2013[J]. Archives of Virology, 2015, 160(4): 1055-1064.
[24] 郭思杰, 刘俊琦, 李玉娟, 等. 猪流行性腹泻及新型疫苗研究进展[J]. 中国畜牧兽医, 2020, 47(9): 2935-2944.
GUO S J, LIU J Q, LI Y J, et al. Research advances on porcine epidemic diarrhea and new vaccine[J]. China Animal Husbandry & Veterinary Medicine, 2020, 47(9): 2935-2944. (in Chinese)
[25] OH J, LEE K W, CHOI H W, et al. Immunogenicity and protective efficacy of recombinant S1 domain of the Porcine epidemic diarrhea virus spike protein[J]. Archives of Virology, 2014, 159(11): 2977-2987.