[1]杨振兴,朱建波,肖 雷,等.中山病多克隆抗体竞争ELISA检测方法的建立[J].中国预防兽医学报,2018,(07):601-606.[doi:0.3969/j.issn.1008-0589.201711023]
 YANG Zhen-xing,ZHU Jian-bo,XIAO Lei,et al.Establishment of a competitive ELISA for detecting antibodies against Chuzan disease virus[J].Chinese journal of preventive veterinary medicine,2018,(07):601-606.[doi:0.3969/j.issn.1008-0589.201711023]
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中山病多克隆抗体竞争ELISA检测方法的建立()
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2018年07
页码:
601-606
栏目:
出版日期:
2018-08-15

文章信息/Info

Title:
Establishment of a competitive ELISA for detecting antibodies against Chuzan disease virus
文章编号:
1008-0589(2018)7-0601-06
作者:
 

杨振兴朱建波肖 雷高 林苗海生何于雯谢佳芮杨 恒*李华春*

 (云南省畜牧兽医科学院 云南省热带亚热带动物病毒病重点实验室,云南 昆明 650224)
Author(s):
 

YANG Zhen-xing ZHU Jian-bo XIAO Lei GAO Lin MIAO Hai-sheng HE Yu-wenXIE Jia-rui YANG Heng* LI Hua-chun*

 (Yunnan Veterinary and Animal Science Institute, Kunming 650224, China)
关键词:
 中山病病毒多克隆抗体竞争 ELISA
Keywords:
 Chuzan disease virus  multi-clone antibody  competitive ELISA
分类号:
S852.65
DOI:
0.3969/j.issn.1008-0589.201711023
文献标志码:
A
摘要:
为建立一种简便、快速的中山病血清抗体监测方法,本研究采用纯化的中山病病毒(CHUV)包被ELISA反应板,兔抗CHUV多克隆抗体作为竞争抗体,建立了检测CHUV抗体的竞争ELISA (C-ELISA)方法。结果显示:抗原最佳包被浓度为4.12 μg/mL,竞争抗体最佳使用浓度为1∶12 000倍。特异性试验显示该C-ELISA方法仅能检测CHUV抗体,其它相关病毒阳性血清检测结果均为阴性。对已知阳性样品的检测结果表明,本研究建立的C-ELISA方法敏感性高于血清中和试验和琼脂扩散试验。分别用C-ELISA、RT-PCR和病毒分离试验对监控动物每周采集的血清和抗凝血样品进行检测,ELISA结果与其它两种方法检测结果对应一致。本研究建立的C-ELISA为CHUV抗体的检测提供了一种快速、方便、可靠的方法。
 
Abstract:
In order to establish a convenient and rapid method for monitoring serum antibody against Chuzan disease virus (CHUV), a competitive ELISA (C-ELISA) was established using purified viral protein as coating antigen and rabbit polyclonal antibody as competitive antibody. The results showed that the optimal concentration of antigen was 4.12μg/mL and the optimal concentration of antibody was 1∶12,000. Specific tests showed that this method specifically detect the antibodies against CHUV. The detection results of known positive samples showed that the sensitivity of C-ELISA established in this study was higher than that of the serum-neutralization test and agar gel immunodiffusion test. The clinical samples collected from monitoring animals were detected by the C-ELISA, RT-PCR and virus isolation, respectively. The results of the C-ELISA were consistent with those of the other two methods. The C-ELISA established in this study provides a rapid, convenient and sensitive method for detecting the antibodies against CHUV.

参考文献/References:

[1]Tateyama S, Yamaguchi R, Uchida K, et al. An outbreak of congenital hydranencephaly and cerebellar hypoplasia among calves in South Kyusyu [J]. Res Vet Sci, 1990, 49(2): 127-131.
[2]于维军. 一种新的环状病毒-Chuzan病毒[J]. 中国兽医杂志,1990,16(11):49-52.
[3]Yamaguchi R, Naitoh Y, Uchida K, et al. Encephalopathy in suckling mice infected with Kasba (Chuzan) virus [J]. J Comp Path, 1999, 120(1): 247-256.
[4]Yamakawa M, Kubo M, Furuuchi S. Molecular analysis of the genome of Chuzan virus, a member of the Palyam serogroup viruses, and its phylogenetic relationships to other orbiviruses [J]. J Gen Virol, 1990, 80(4): 937-941.
[5]Yamakawa M, Furuuchi S, Minobe Y. Molecular characterization of double-stranded RNA segments encoding the major capsid proteins of a Palyam serogroup orbivirus that caused an epizootic of congenital abnormalities in cattle [J]. J Gen Virol, 1999, 80(1): 205-208.
[6]Miura Y, Goto Y, Kubo M, et al. Isolation of Chuzan virus, a new member of the Palyam subgroup of the genus Orbivirus, from cattle and Culicoides oxystoma in Japan [J]. Am J Vet Res, 1988, 49: 2022-2025.
[7]张义爽,王芳,武瑞,等. 我国中山病的血清学调查[J]. 中国预防兽医学报,2017,39(1):1-4.
[8]Kato T, Shirafuji H, Tanaka S, et al. Bovine arboviruses in culicoides biting midges and sentinel cattle in southern Japan from 2003 to 2013 [J]. Transbound Emerg Dis, 2016, 63(6): e160-e172.
[9]Lim S I, Kweon C H, Tark D S, et al. Sero-survey on Aino, Akabane, Chuzan, bovine ephemeral fever and Japanese encephalitis virus of cattle and swine in Korea [J]. J Vet Sci, 2007, 8(1): 45-49.
[10]刘焕章,昊东来,胡守萍. 赤羽病、中山病和茨城病流行病学调查初报[J]. 中国奶牛,2005,(5):46-47.
[11]Yamaguchi R, Tanimoto N, Tateyama S, et al. Immunohistochemical study of age-dependent brain lesions in mice infected intracerebrally with Kasba (Chuzan) Virus [J]. J Comp Path, 2001, 124: 36-45.
[12]Yang Heng, Xiao Lei, Meng Jin-xin, et al. Complete genome sequence of a Chuzan virus strain isolated for the first time in mainland China [J]. Arch Virol, 2016, 161(4): 1073-1077.
[13]Wang Fang, Lin Jun, Chang Ji-tao, et al. Isolation, complete genome sequencing, and phylogenetic analysis of the first Chuzan virus in China [J]. Virus Genes, 2016, 52(1): 138-141.
[14]李雪平,邓国华,高玉伟,等. 检测流感病毒H5亚型抗体竞争ELISA方法的建立及初步应用[J]. 中国预防兽医学报,2008,30(9):716-720.
[15]Goto Y, Miura Y, Kono Y. Serologic evidence for the etiologic of Chuzan virus in an epizootic of congenital abnormalities with hydranencephaly-cerebellar hypoplasia syndrome of calves in Japan [J]. Am J Vet Res, 1988, 49: 2026-2029.
(本文编辑:李 爽

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更新日期/Last Update: 2018-08-15