2013—2022年云南地区猪肠病毒G型的分子流行病学调查

doi:10.16431/j.cnki.1671-7236.2023.12.022

中国畜牧兽医 ›› 2023, Vol. 50 ›› Issue (12): 5000-5012.doi: 10.16431/j.cnki.1671-7236.2023.12.022

2013—2022年云南地区猪肠病毒G型的分子流行病学调查

王位¹, 朱沛², 李占鸿², 薛晓岩¹, 杨钦鸿¹, 李雪³, 李素华¹, 姚俊²

1. 西南林业大学生命科学学院, 昆明 650224;
2. 云南畜牧兽医科学院, 云南省热带亚热带动物病毒病重点实验室, 昆明 650224;
3. 云南农业大学动物医学院, 昆明 650500

收稿日期:2023-04-07 出版日期:2023-12-05 发布日期:2023-11-28
作者简介:王位,E-mail:1584102842@qq.com;朱沛,E-mail:zpcau@sina.com。

Molecular Epidemiological Survey of Porcine Enterovirus Type G in Yunnan from 2013 to 2022

WANG Wei¹, ZHU Pei², LI Zhanhong², XUE Xiaoyan¹, YANG Qinhong¹, LI Xue³, LI Suhua¹, YAO Jun²

1. School of Life Sciences, Southwest Forestry University, Kunming 650224, China;
2. Key Laboratory of Tropical Subtropical Animal Virus Diseases, Yunnan Academy of Animal Husbandry and Veterinary Science, Kunming 650224, China;
3. College of Veterinary Animal Medicine, Yunnan Agricultural University, Kunming 650500, China

Received:2023-04-07 Online:2023-12-05 Published:2023-11-28
Contact: 云南省农业重大科技专项计划"云南省猪重要疫病防控技术体系构建及应用"(202102AE090007);云南省财政专项-畜禽种质创新及标准化养殖技术推广课题"牛结节性皮肤病防控技术研究与示范"(云财农【2022】5号);乡村振兴科技专项-科技特派团(202304BI090003) E-mail:li.suhua@hotmail;yaojun_joshua@hotmail.com
Supported by:
The project was supported by the National Key Research and Development Program of China (2019YFC1905301);National Natural Science Foundation of China (22078115,21776108,21690083,22008078).

摘要/Abstract

摘要： 【目的】了解肠病毒G型(Enterovirus G,EV-G)在云南地区猪场中的流行状况及其分子特征,为云南地区EV-G的防控提供理论依据。【方法】根据EV-G的5'-非翻译区(UTR)保守区域设计引物,RT-PCR扩增监测2013－2022年云南地区猪临床腹泻样品的EV-G阳性率,从阳性样品来源的区域、猪群结构及类型分析EV-G在云南地区的感染情况。利用EV-G VP1基因巢式PCR引物,通过RT-PCR扩增EV-G VP1基因全序列;采用MegAlign软件分析EV-G云南株的VP1序列与EV-G的20个基因型标准株的相似性及关键氨基酸突变,采用Mega 7.0软件构建VP1基因的遗传进化树,并通过Protean软件的Jameson-Wolf分析法对VP1蛋白进行抗原指数分析。【结果】云南地区2013－2022年收集的1 941份临床样品EV-G总阳性率为19.42%(377/1 941),其中大理(54.54%)、玉溪(45.45%)、红河(43.24%)猪群的EV-G感染阳性率明显高于云南其他地区;乳猪(25.19%)和保育猪(24.68%)2个年龄组的猪群阳性感染率高于其他猪群;肛拭子阳性样品的检出率(53.58%)高于肠道内容物(29.71%)及粪便(16.71%)样品。基于20株EV-G代表株VP1基因序列相似性分析及遗传进化表明,17个云南株为EV-G1型,2株为EV-G6型,1株为EV-G17型。17个EV-G1基因型云南分离株VP1蛋白在第125－150、170－190位氨基酸处抗原指数活跃度显著增强,此区域同时存在多个亲水性氨基酸位点的突变,如P¹²⁷S、P¹⁴³S、G¹⁴⁶R、P¹⁸⁸S。【结论】EV-G1、EV-G6和EV-G17等多种基因型在云南地区猪群中流行,以EV-G1型流行为主。EV-G1基因型云南分离株的VP1蛋白在第125－150、170－190位氨基酸处存在显著的抗原活跃度,可能介导EV-G1基因型毒株在云南的感染和流行。

关键词: 肠病毒G型(EV-G); VP1; 分子流行病学; 基因特征

Abstract: 【Objective】 This study was aimed to understand the prevalence and molecular characteristics of Enterovirus G (EV-G) in pig farms in Yunnan,so as to provide the theoretical basis for the prevention and control of EV-G in Yunnan.【Method】 RT-PCR basing on the primers from the highly conserved region of the 5'-untranslated region (UTR) of EV-G was used for the detection of EV-G-positive samples from clinical diarrhea samples of pigs collected in Yunnan from 2013 to 2022.The farm distribution,swine age composition and the type of positive samples were analyzed to determine the infection characteristics of EV-G in Yunnan.The complete sequence of EV-G VP1 gene was amplified using nested PCR specific primers.MegAlign software was used to analyze the similarity and key amino acid mutations of VP1 between Yunnan strains and the reference strains of 20 EV-G genotypes.A genetic evolutionary tree of VP1 gene was constructed by Mega 7.0 software.Additionally,the antigenic index of VP1 protein was analyzed using the Jameson-Wolf method in Protean software.【Result】 A total positive rate of 19.42% (377/1 941) for EV-G was observed in the clinical samples collected in Yunnan from 2013-2022.Among them,the EV-G infection rates in pig populations from Dali (54.54%),Yuxi (45.45%) and Honghe (43.24%) were significantly higher than that in other regions of Yunnan.The positive infection rates in suckling piglets (25.19%) and nursery pigs (24.68%) were higher than that in other pig age groups.The detection rate of positive cloacal swab (53.58%) was higher than that of intestinal contents samples (29.71%) and fecal samples (16.71%).According to the analysis of the similarity and genetic evolution of VP1 gene of EV-G showed that 17 strains from 20 Yunnan representatives belonged to EV-G1 genotype,2 strains belonged to EV-G6 genotype,and 1 strain belonged to EV-G17 genotype.VP1 protein of 17 Yunnan isolates of EV-G1 genotype exhibited significantly enhanced antigenic activity at positions 125-150 and 170-190,where multiple hydrophilic amino acid mutations were observed (P¹²⁷S,P¹⁴³S,G¹⁴⁶R and P¹⁸⁸S).【Conclusion】 Multiple genotypes such as EV-G1,EV-G6 and EV-G17 were prevalent in pig populations in Yunnan,with the EV-G1 type being the main genotype.VP1 protein of EV-G1 genotype Yunnan isolate exhibited significant antigenic activity at 125-150 and 170-190 amino acids,which might mediate the infection and epidemic of EV-G1 genotype strains in Yunnan.

Key words: Enterovirus G (EV-G); VP1; molecular epidemiology; genetic characteristics

中图分类号:

S852.65⁺1

王位, 朱沛, 李占鸿, 薛晓岩, 杨钦鸿, 李雪, 李素华, 姚俊. 2013—2022年云南地区猪肠病毒G型的分子流行病学调查[J]. 中国畜牧兽医, 2023, 50(12): 5000-5012.

WANG Wei, ZHU Pei, LI Zhanhong, XUE Xiaoyan, YANG Qinhong, LI Xue, LI Suhua, YAO Jun. Molecular Epidemiological Survey of Porcine Enterovirus Type G in Yunnan from 2013 to 2022[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(12): 5000-5012.

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2013—2022年云南地区猪肠病毒G型的分子流行病学调查

Molecular Epidemiological Survey of Porcine Enterovirus Type G in Yunnan from 2013 to 2022

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