2011—2016年浙江省猪流行性腹泻病毒S基因与ORF3基因遗传进化分析

doi:10.3969/j.issn.1004-1524.2017.03.05

浙江农业学报 ›› 2017, Vol. 29 ›› Issue (3): 380-388.DOI: 10.3969/j.issn.1004-1524.2017.03.05

2011—2016年浙江省猪流行性腹泻病毒S基因与ORF3基因遗传进化分析

吴旺霞^{1, 2}, 王一成^{2, *}, 吴润¹, 袁秀芳², 徐丽华², 李军星², 苏菲²

1.甘肃农业大学动物医学院,甘肃兰州 730000;
2.浙江省农业科学院畜牧兽医研究所,浙江杭州 310021

收稿日期:2016-08-17 出版日期:2017-03-20 发布日期:2017-03-31
通讯作者: 王一成,E-mail:95711@sina.com
作者简介:吴旺霞(1991—),女,甘肃白银人,硕士研究生,研究方向为兽医微生物与免疫学。E-mail:1174478344@qq.com
基金资助:
公益技术研究农业项目(2015C32028,2016C32072)

Genetic analysis of S and ORF3 gene in Porcihe epidemic diarrhea virus during 2011—2016 in Zhejiang Province

WU Wangxia1^{1, 2}, WANG Yicheng2^{2, *}, WU Run¹, YUAN Xiufang², XU Lihua², LI Junxing², SU Fei²

1. College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China;
2. Institute of Animal Husbandry and Veterinary Science, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China

Received:2016-08-17 Online:2017-03-20 Published:2017-03-31

摘要/Abstract

摘要： 为调查近年来浙江省猪流行性腹泻病毒流行毒株与疫苗株的S基因和ORF3基因遗传变异情况,2011—2016年,从浙江省各地区采集疑似猪流行性腹泻病料并提取其RNA,用3对特异性引物对S基因和ORF3基因进行RT-PCR扩增、克隆及测序,并对其序列变化、遗传进化情况和抗原位点进行分析。结果表明:与国内疫苗株CV777相比,12个样品株的S基因存在15个核苷酸插入和6个核苷酸缺失,导致氨基酸序列存在5个氨基酸插入(⁵⁸QGVN⁶¹和^l40N)和2个氨基酸缺失(¹⁶³DI¹⁶⁴),且在主要突变区S1区的2个中和表位(499-638和764-771 aa)有8处氨基酸突变;样品株的ORF3基因有7处氨基酸变异。遗传进化分析结果显示,12株分离株的S基因与亚洲主要疫苗株(国内疫苗株CV777和韩国弱毒疫苗株DR13)的同源性仅为93.3%~94.9%,而与2011—2016年中国流行株(BJ-2011和HuN)的同源性高达97.8%~99.9%,表明近年来猪流行性腹泻病毒呈现较大程度的变异,可为研制更加有效的疫苗奠定基础。

关键词: 猪流行性腹泻病毒, S基因, ORF3基因, 序列特点, 基因进化分析

Abstract: In order to investigate recent genetic variations of S and ORF3 genes in Porcine epidemic diarrhea virus(PEDV) epidemic and vaccine strains in Zhejiang Province, virus RNA was extracted from suspected PEDV samples which were collected from various areas in Zhejiang Province since 2011 to 2016, then RT-PCR, cloning and sequencing on S and ORF3 genes were performed using three pairs of specific primers to analyze the sequence variation, genetic evolution and antigen sites. The results showed that as compared to vaccine strain CV777, 15 nucleotides were inserted and 6 nucleotides were absent in the S genes of 12 epidemic strains, leading to the insertion of 5 amino acids (⁵⁸QGVN⁶¹and¹⁴⁰N) and deletion of 2 amino acids (¹⁶³DI¹⁶⁴) relatively. Moreover, in epidemic strains, totally 8 amino acid mutations were found in two neutralizing epitopes (499-638 and 764-771 aa) on the major mutation region S1; 7 amino acids were variated in ORF3 gene. According to the analysis results of genetic evolution, these 12 epidemic strains shared 97.8%-99.9% homology with China epidemic strains BJ-2011 and HuN in S gene, but only 93.3%-94.9% homology with vaccine strains (CV777 and Korean attenuated strain DR13), indicating that huge variations and fast evolution had emerged in PEDV recently. This study could lay a foundation for development of effective vaccines and therapies.

Key words: Porcine epidemic diarrhea virus, S gene, ORF3 gene, sequence characteristics, genetic evolution analysis

中图分类号:

S858.28

吴旺霞, 王一成, 吴润, 袁秀芳, 徐丽华, 李军星, 苏菲. 2011—2016年浙江省猪流行性腹泻病毒S基因与ORF3基因遗传进化分析[J]. 浙江农业学报, 2017, 29(3): 380-388.

WU Wangxia1, WANG Yicheng2, WU Run, YUAN Xiufang, XU Lihua, LI Junxing, SU Fei. Genetic analysis of S and ORF3 gene in Porcihe epidemic diarrhea virus during 2011—2016 in Zhejiang Province[J]. , 2017, 29(3): 380-388.

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2011—2016年浙江省猪流行性腹泻病毒S基因与ORF3基因遗传进化分析

Genetic analysis of S and ORF3 gene in Porcihe epidemic diarrhea virus during 2011—2016 in Zhejiang Province

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