乳胶凝集试验方法在检测鸭坦布苏病毒抗体中的应用

万春和; 傅秋玲; 陈珍; 傅光华; 施少华; 程龙飞; 陈红梅; 黄瑜

doi:10.19303/j.issn.1008-0384.2013.12.002

乳胶凝集试验方法在检测鸭坦布苏病毒抗体中的应用

doi: 10.19303/j.issn.1008-0384.2013.12.002

福建省农业科学院畜牧兽医研究所/福建省畜鸭疫病防治工程技术研究中心,福建福州,350013

基金项目:

福建省财政专项——福建省农业科学院创新团队建设项目

福建省自然科学基金项目 (2012J01112)

现代农业产业体系建设专项资金 (CARS-43)

详细信息

作者简介:
万春和 (1982-), 男, 在读博士生, 助理研究员, 主要从事水禽病毒分子生物学研究;黄瑜 (1966-), 男, 博士, 研究员, 主要从事动物传染病研究 (E-mail:huangyu_815@163.com)

中图分类号: S858.32
计量
- 文章访问数: 135
- HTML全文浏览量: 26
- PDF下载量: 3
- 被引次数: 0
出版历程
- 收稿日期: 2013-11-03
- 刊出日期: 2013-12-18

Rapid Detection of Duck Tembusu Virus Antibody by Latex Agglutination Test

Institute of Animal Husbandry and Veterinary Medicine,Fujian Academy of Agricultural Sciences/Fujian Animal Disease Control Technology Development Center

摘要

摘要: 以经超速和密度梯度离心浓缩纯化的鸭坦布苏病毒抗原进行方阵滴定, 筛选出致敏乳胶的最佳条件, 并制备成鸭坦布苏病毒乳胶凝集试验 (LAT) 用抗原, 与特异性的鸭坦布苏病毒阳性血清反应出现肉眼可见的凝集颗粒, 进而建立检测鸭坦布苏病毒抗体的乳胶凝集试验方法。经试验测定该致敏的抗原与鸭流感病毒阳性血清、鸭副粘病毒阳性血清、鸭肝炎病毒阳性血清、鸭产蛋综合症病毒阳性血清和鸭呼肠孤病毒阳性血清均不出现凝集现象, 表明具有良好的特异性。以建立的LAT方法和间接ELISA同时分别对80份鸭血清进行检测, 结果 LAT方法检出阳性58份、阴性22份, 而间接ELISA方法检出阳性68份、阴性12份, 两者阳性符合率达85.3%。以上结果表明, 本方法具有简便、快速、特异等优点, 可用于临床样品的快速诊断和血清流行病学调查。
- 鸭坦布苏病毒 /
- 乳胶凝集试验 /
- 抗体检测
Abstract: Duck tembusu viruses were cultivated and concentrated by ultracentrifugation and density-gradient centrifugation to determine the best suitable conditions for sensitized latex and prepare antigen for the latex agglutination test (LAT) by titration.The agglutination particles could be observed clearly by naked eyes after antigen reaction with positive duck tembusu virus serum.The sensitized antigen had no cross-reaction with avian influenza virus, avian paramyxovirus type 1, duck hepatitis virus type 1, egg drop syndrome virus, and duck reovirus.Of 80duck serums, 58and 68 were detected to be positive by the established LAT and ELISA, respectively;the positive rates for both were above 85.3% with no statistical difference.The results revealed that the established LAT was simple, quick and specific for the rapid diagnose and survey of duck tembusu virus antibody.
- duck tembusu virus /
- latex agglutination test /
- antibody detection

HTML全文

参考文献(23)

[1]	万春和, 施少华, 程龙飞, 等.一种引起种 (蛋) 鸭产蛋骤降新病毒的分离与初步鉴定[J].福建农业学报, 2010, 25 (6) :663-666.
[2]	曹贞贞, 张存, 黄瑜, 等.鸭出血性卵巢炎的初步研究[J].中国兽医杂志, 2010, 46 (12) :3-6.
[3]	SU J, LI S, HU X, et al.Duck egg-drop syndrome caused by BYD virus, a new Tembusu-realated Flavivirus[J].PLoS one6 (3) :e18106.
[4]	CAO Z, ZHANG C, LIU Y, et al.Tembusu virus in ducks, China[J].Emerg Infect Dis, 2011, 17 (10) :1873-1875.
[5]	YAN P, ZHAO Y, ZHANG X, et al.An infectious disease of ducks caused by a newly emerged Tembusu virus strain in mainland China[J].Virology, 2011, 417 (1) :1-8.
[6]	LIU M, LIU C, LI G, et al.Complete genomic sequence of duck flavivirus from china[J].J Virol, 2012, 86 (6) :3398-3399.
[7]	WAN C, HUANG Y, FU G, et al.Complete genome sequence of avian tembusu-related virus strain WR isolated from White Kaiya ducks in Fujian, China[J].J Virol, 2012, 86 (19) :10912.
[8]	刘利芳.乳胶凝集试验、间接ELISA诊断鸭病毒性肝炎方法的建立[D].福州:福建农林大学, 2008.
[9]	万春和, 黄瑜.坦布苏病毒致病特性及其流行病学[J].动物医学进展, 2011, 32 (12) :116-118.
[10]	LIU P, LU H, LI S, et al.Duck egg drop syndrome virus:an emerging Tembusu-related flavivirus in China[J].Sci China Life Sci, 2013, 56 (8) :701-710.
[11]	傅光华, 黄瑜, 施少华, 等.鸡黄病毒的分离与初步鉴定[J].福建畜牧兽医, 2011, 33 (3) :1-2.
[12]	陈仕龙, 陈少莺, 王劭, 等.一种引起蛋鸡产蛋下降的新型黄病毒的分离与初步鉴定[J].福建农业学报, 2011, 26 (2) :170-174.
[13]	黄欣梅, 李银, 赵冬敏, 等.新型鹅黄病毒JS804毒株的分离与鉴定[J].江苏农业学报, 2011, 27 (2) :354-360.
[14]	施少华, 傅光华, 万春和, 等.鹅源坦布苏病毒的分离与初步鉴定[J].中国兽医杂志, 2012, 48 (12) :37-40.
[15]	TANG Y, GAO X, DIAO Y, et al.Tembusu virus in human, China[J].Transbound Emerg Dis, 2013, 60 (3) :193-196.
[16]	万春和, 施少华, 傅光华, 等.鸭黄病毒油乳剂灭活疫苗研制及免疫效果测定[J].养禽与禽病防治, 2011, 10:20-22.
[17]	YU K, SHENG Z Z, HUANG B, et al.Structural, antigenic, and evolutionary characterizations of the envelope protein of newly emerging Duck Tembusu Virus[J].PLoS One, 2013, 8 (8) :e71319.
[18]	万春和, 施少华, 程龙飞, 等.鸭出血性卵巢炎病毒RTPCR检测方法的建立[J].福建农业学报, 2011, 26 (1) :10-12.
[19]	万春和, 朱海侠, 施少华, 等.鸭坦布苏病毒SYBR Green I实时荧光定量检测方法的建立[J], 中国兽医学报, 2013, 33 (7) :973-978.
[20]	YAN L, YAN P, ZHOU J, et al.Establishing a TaqManbased real-time PCR assay for the rapid detection and quantification of the newly emerged duck Tembusu virus[J].Virol J, 2011, 8:464.
[21]	WANG Y, YUAN X, LI Y, et al.Rapid detection of newly isolated Tembusu-related Flavivirus by reverse-transcription loop-mediated isothermal amplification assay[J].Virol J, 2011, 8:553.
[22]	YIN X, LR, CHEN X, et al.Detection of specific antibodies against tembusu virus in ducks by use of an E protein-based enzyme-linked immunosorbent assay[J].J Clin Microbiol, 2013, 51 (7) :2400-2402.
[23]	LI X, LI G, TENG Q, et al.Development of a blocking ELISA for detection of serum neutralizing antibodies against newly emerged duck Tembusu virus[J].PLoS One, 2012, 7 (12) :e53026.