禽戊型肝炎病毒组织切片免疫荧光检测方法的建立及应用

doi:10.11843/j.issn.0366-6964.2014.11.016

畜牧兽医学报 ›› 2014, Vol. 45 ›› Issue (11): 1851-1857.doi: 10.11843/j.issn.0366-6964.2014.11.016

禽戊型肝炎病毒组织切片免疫荧光检测方法的建立及应用

陈瑞爱^1,2,3*，李延鹏³，同戈¹，郭凯¹，刘正伟¹

（1.华南农业大学兽医学院，广州 510642；2.农业部动物疫病防控生物技术与制品创制重点实验室，肇庆 526238；3.广东省兽用生物制品生物技术研究与应用企业重点实验室，肇庆 526238）

收稿日期:2014-03-31 出版日期:2014-11-23 发布日期:2014-11-23
通讯作者: 陈瑞爱
作者简介:陈瑞爱(1970-)，女，广东新兴人，教授，主要从事免疫抑制病研究，E-mail:chenruiaidhn@126.com
基金资助:
重大动物疫病新型疫苗的关键技术研究与产业化（2011A090200117）

Establishment and Application of Tissue Section Immunofluorescence Method for Detecting Avian Hepatitis E Virus

CHEN Rui-ai^1,2,3*，LI Yan-peng³，TONG Ge¹，GUO Kai¹，LIU Zheng-wei¹

（1.College of Veterinary Medicine，South China Agricultural University，Guangzhou 510642，China；2.Key Laboratory of Biotechnology and Bioproducts Development for Animal Epidemic Prevention of Ministry of Agriculture，P.R.China，Zhaoqing 526238，China；3.Guangdong Provincial Key Laboratory of Biotechnology R & D of Veterinary Biological Products，Zhaoqing 526238，China）

Received:2014-03-31 Online:2014-11-23 Published:2014-11-23
Supported by:

摘要/Abstract

摘要：

为建立对禽戊型肝炎病毒（HEV）鉴别诊断的组织切片免疫荧光方法，制备禽HEV ORF2蛋白的单因子血清，对感染HEV、禽白血病病毒（ALV）、禽网状内皮组织增生症病毒（REV）鸡的病变肝组织切片进行间接免疫荧光检测。结果显示制备的禽HEV ORF2蛋白的单因子血清具有良好的特异性，利用组织切片免疫荧光方法能够特异地检测禽HEV的感染，从而有效地区分HEV与易混淆的ALV、REV的感染。建立的鉴别诊断禽HEV的组织切片免疫荧光方法，可以有效地诊断禽HEV感染，给实验室诊断带来极大的方便。

Abstract:

Immunofluorescence staining method on paraffin-embedded liver sections was established to differentially detect avian HEV in this experiment.Mono-specific serum of HEV ORF2 protein were prepared and used for detecting tissue section of lesion liver of chickens infected with HEV，avian leukosis virus(ALV)，reticuloendotheliosis virus (REV)，respectively.Results showed that the prepared mono-specific serum of HEV ORF2 protein showed good specificity.The established immunofluorescence method could detect avian HEV infection and was effective to distinguish HEV from ALV or REV infection.Our results revealed that the novel immunofluorescence method brings great convenience for laboratory diagnosis.

中图分类号:

陈瑞爱，李延鹏，同戈，郭凯，刘正伟. 禽戊型肝炎病毒组织切片免疫荧光检测方法的建立及应用[J]. 畜牧兽医学报, 2014, 45(11): 1851-1857.

CHEN Rui-ai，LI Yan-peng，TONG Ge，GUO Kai，LIU Zheng-wei. Establishment and Application of Tissue Section Immunofluorescence Method for Detecting Avian Hepatitis E Virus[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(11): 1851-1857.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.xmsyxb.com/CN/10.11843/j.issn.0366-6964.2014.11.016

https://www.xmsyxb.com/CN/Y2014/V45/I11/1851

参考文献

［1］HAQSHENAS G，SHIVAPRASAD H L，WOOLCOCK P R，et al.Genetic identification and characterization of a novel virus related to human hepatitis E virus from chickens with hepatitis-splenomegaly syndrome in the United States ［J］.J Gen Virol，2001，82(10)：2449-2462.
［2］PAYNE C J，ELLIS T M，PLANT S L，et al.Sequence data suggests big liver and spleen disease virus (BLSV) is genetically related to hepatitis E virus ［J］.Vet Microbiol, 1999，68(1-2)：119-125.
［3］HAQSHENAS G，HUANG F F，FENAUX M，et al.The putative capsid protein of the newly identified avian hepatitis E virus shares antigenic epitopes with that of swine and human hepatitis E viruses and chicken big liver and spleen disease virus ［J］.J Gen Virol，2002，83(9)：2201-2209.
［4］BILLAM P，HUANG F F，SUN Z F，et al.Systematic pathogenesis and replication of avian hepatitis E virus in specific-pathogen-free adult chickens ［J］.J Virol，2005，79(6)：3429-3437.
［5］HUANG F F，SUN Z F，EMERSON S U，et al.Determination and analysis of the complete genomic sequence of avian hepatitis E virus (avian HEV) and attempts to infect rhesus monkeys with avian HEV ［J］.J Gen Virol，2004，85(6)：1609-1618.
［6］HUANG F F，HAQSHENAS G，SHIVAPRASAD H L，et al.Heterogeneity and seroprevalence of a newly identified avian hepatitis e virus from chickens in the United States ［J］.J Clin Microbiol，2002，40(11)：4197-4202.
［7］VITRAL C L，PINTO M A，LEWIS-XIMENEZ L L，et al.Serological evidence of hepatitis E virus infection in different animal species from the Southeast of Brazil ［J］.Mem Inst Oswaldo Cruz，2005，100(2)：117-122.
［8］梁久红，孟继鸿.人、猪、禽戊型肝炎病毒血清学关系的研究［J］.中华微生物学和免疫学杂志，2007，27(7)：581-586.
［9］GUO H，ZHOU E M，SUN Z F，et al.Identification of B-cell epitopes in the capsid protein of avian hepatitis E virus (avian HEV) that are common to human and swine HEVs or unique to avian HEV ［J］.J Gen Virol，2006，87(1)：217-223.
［10］张璐，周恩民，崔治中，等.禽戊型肝炎血清学调查以及与肝脾肿大综合征关系的研究［J］.中国家禽，2008，30(20)：22-24.
［11］ZHAO Q，SUN Y，ZHAO J，et al.Development and application of an indirect ELISA for detection of antibodies against avian hepatitis E virus ［J］.J Virol Methods，2013，187(1)：32-36.
［12］SUN Z F，LARSEN C T，DUNLOP A，et al.Genetic identification of avian hepatitis E virus (HEV) from healthy chicken flocks and characterization of the capsid gene of 14 avian HEV isolates from chickens with hepatitis-splenomegaly syndrome in different geographical regions of the United States［J］.J Gen Virol，2004，85(3)：693-700.
［13］AGUNOS A C，YOO D，YOUSSEF S A，et al.Avian hepatitis E virus in an outbreak of hepatitis-splenomegaly syndrome and fatty liver haemorrhage syndrome in two flaxseed-fed layer flocks in Ontario［J］.Avian Pathol，2006，35(5)：404-412.
［14］BILIC I，JASKULSKA B，BASIC A.Sequence analysis and comparison of avian hepatitis E viruses from Australia and Europe indicate the existence of different genotypes ［J］.J Gen Virol，2009，90(4)：863-873.
［15］MAREK A，BILIC I，PROKOFIEVA I，et al.Phylogenetic analysis of avian hepatitis E virus samples from European and Australian chicken flocks supports the existence of a different genus within the Hepeviridae comprising at least three different genotypes ［J］.Vet Microbiol，2010，145(1-2)：54-61.
［16］ZHAO Q，ZHOU E M，DONG S W，et al.Analysis of avian hepatitis E virus from chickens，China ［J］.Emerg Infect Dis，2010，16(9)：1469-1472.
［17］SUN Z F，LARSEN C T，HUANG F F，et al.Generation and infectivity titration of an infectious stock of avian hepatitis E virus (HEV) in chickens and cross-species infection of turkeys with avian HEV ［J］.J Clin Microbiol，2004，42(6)：2658-2662.
［18］PAYNE C J，PLANT S L，ELLIS T M，et al.The detection of the big liver and spleen agent in infected tissues via intravenous chick embryo inoculation ［J］.Avian Pathol，1993，22(2)：245-256.
［19］GUO H，ZHOU E M，SUN Z F，et al.Protection of chickens against avian hepatitis E virus (avian HEV) infection by immunization with recombinant avian HEV capsid protein ［J］.Vaccine，2007，25(15)：2892-2899.
［20］RIDDELL M A，LI F，ANDERSON D A，et al.Identification of immunodominant and conformational epitopes in the capsid protein of hepatitis E virus by using monoclonal antibodies ［J］.J Virol，2000，74(17)：8011-8017.
［21］DONG S，ZHAO Q，LU M，et al.Analysis of epitopes in the capsid protein of avian hepatitis E virus by using monoclonal antibodies ［J］.J Virol Methods，2011，171(2)：374-380.

[1]	刘健, 白艺兰, 李鑫, 桂亚萍, 鞠厚斌, 龚国华, 夏炉明, 陈伟锋, 朱晓英, 常晓静, 李增强, 唐聪圣, 王建, 赵洪进. 猫杯状病毒分离株VP1基因序列与致病性分析[J]. 畜牧兽医学报, 2022, 53(10): 3530-3539.
[2]	郭振华, 阮海宇, 耿瑞, 陈鑫鑫, 乔松林, 邓瑞广, 张改平. 1株类NADC30与类JXA1重组的猪繁殖与呼吸综合征病毒基因组遗传特征分析[J]. 畜牧兽医学报, 2020, 51(7): 1677-1687.

禽戊型肝炎病毒组织切片免疫荧光检测方法的建立及应用

Establishment and Application of Tissue Section Immunofluorescence Method for Detecting Avian Hepatitis E Virus

RichHTML

PDF

可视化

被引次数

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 2

编辑推荐

Metrics

本文评价