畜牧兽医学报2018，Vol.49Issue(4)：761-770,10.DOI:10.11843/j.issn.0366-6964.2018.04.013
2012-2016年中国南方地区帕利亚姆血清群病毒的分离与序列特征分析
Sequence Analysis of Palyam Serogroup Virus Isolated in South China from 2012 to 2016
摘要
Abstract
The aim of this study was to isolate and identify the Palyam serogroup virus (PALV) prevalent in China from the sentinel herds and to clarify the relationship between the native strains and the foreign strains.For virus isolation,PALV-positive blood samples were collected from sentinel herds and blind passed on BHK-21 cells.Specific primers targeting Seg-2 and Seg7 of PALV were designed for identification and sequence analysis of the isolated virus by RT-PCR amplification.Genomic RNA of the isolated virus was extracted from the infected cells and determined by high resolution agarose gel electrophoresis.Cross-reactivity and cross-neutralizing activity of positive sera from different serotypes of PALV were analyzed by immunofluorescence and serum neutralization test.The results were as follows:During 2012 to 2016,a total of 19 strains of PALV were isolated form Yunnan,Guangxi and Guangdong provinces.Seg-2 and Seg7 sequencing and phylogenetic analysis showed that the Chinese strains of PALV belonged to Chuzan virus (CHUV),D'Aguilar virus (DAV) and Bunyip Creek virus (BCV) and showed closest genetic relationship with the Japanese strains,but showed relative far relationship with the Australian and African strains.High resolution agarose gel electrophoresis showed that the genomic dsRNA of different serotypes of representative PALV strains presenting "3-3-4" banding characteristics.Immunofluorescence results showed that BHK-21 cells infected with CHUV,DAV or BCV presented specific fluorescence when using polyclonal antibody against CHUV as detection antibodies.Serological neutralization tests showed that there was no cross-neutralization protection between the positive sera of CHUV,DAV and BCV viruses.This study reported the molecular characteristics of PALV isolated in China from 2012 to 2016.It's the first time,we reported the DAV and BCV isolated in China.These results will be helpful to better understand the distribution and genetic characteristics of Chinese PALV and provide basis for development of diagnosis reagent and carried out epidemiological and pathogenic research on PALV.关键词
帕利亚姆血清群病毒/病毒分离/病毒鉴定/血清型/序列分析/交叉中和保护Key words
Palyam serogroup virus/virus isolation/virus identification/serotype/sequence analysis/cross-neutralization protection分类
农业科技引用本文复制引用
杨恒,李华春,肖雷,李占鸿,孟锦昕,杨振兴,吕敏娜,林栩慧,廖德芳,牛保生..2012-2016年中国南方地区帕利亚姆血清群病毒的分离与序列特征分析[J].畜牧兽医学报,2018,49(4):761-770,10.基金项目
国家重点研发计划(2016YFD0500908) （2016YFD0500908）
公益性行业(农业)科研专项(201303035) （农业）
国家自然科学基金(31360621 （）
31760744) （）
