鲤疱疹病毒Ⅱ型ORF72单克隆抗体的制备及应用

doi:10.16378/j.cnki.1003-1111.23009

Abstract
Figure/Table
References
Related Citation (15)

Download: PDF (6209 KB) HTML (1 KB)
Export: BibTeX | EndNote (RIS)

Abstract In order to establish an immune detection method based on antigen levels, the nucleocapsid protein ORF72 of cyprinid herpesvirus Ⅱ (CyHV-2) was prepared by prokaryotic expression method. The purified ORF72 protein was immunized to BALB/c mice, and the anti-ORF72 monoclonal antibodies designated as 1B3, 4B1, 4C2, 5F4 and 5H9 were finally screened by hybridoma technology. Western blot analysis showed that the monoclonal antibodies specifically recognized recombinant protein ORF72 in vitro. The monoclonal antibody 4C2 detected the virus infection tissues of gibel crucian carp Carassius auratus gibelio by indirect immun of luorescence assay (IFA) only. IFA of the CyHV-2 replication on RyuF-2 detection with 4C2 showed that the positive signals were detected 36 hours after the virus infection, and the expression level of ORF72 protein was increased in the cell with time. Monoclonal antibody 4C2 contributes to research the replication mechanism of CyHV-2 in vitro and to establishment of the immune detection method for diseased fish.

Key words： Carassius auratus gibelio cyprinid herpesvirus 2 ORF72 monoclonal antibody indirect immunofluorescence assay (IFA)

Received: 12 February 2023

S917.1

	Service

	E-mail this article
	Add to my bookshelf
	Add to citation manager
	E-mail Alert
	RSS
	Articles by authors
	XU Chaonan
	SUN Yuyu
	WANG Jia
	GUO Baoqin
	WEI Chang
	LI Qiang

Cite this article:

XU Chaonan,SUN Yuyu,WANG Jia, et al. Preparation and Application of Monoclonal Antibodies against ORF72 of Cyprinid Herpesvirus 2[J]. Fisheries Science, 2024, 43(4): 598-605.

URL:

https://www.shchkx.com/EN/10.16378/j.cnki.1003-1111.23009 OR https://www.shchkx.com/EN/Y2024/V43/I4/598

[1] JIANG N, YUAN D, ZHANG M, et al. Diagnostic case report:disease outbreak induced by CyHV-2 in goldfish in China[J]. Aquaculture,2020,523:735156.
[2] 袁锐,陈静,刘训猛,等.鲤疱疹病毒2型研究进展[J].水产学杂志,2019,32(1):38-45.
[3] JEFFERY K R, BATEMAN K, BAYLEY A, et al. Isolation of a cyprinid herpesvirus 2 from goldfish, Carassius auratus (L. ), in the UK[J]. Journal of Fish Diseases,2007,30(11):649-656.
[4] GOODWIN A E, KHOO L, LAPATRA S E, et al. Goldfish hematopoietic necrosis herpesvirus (cyprinid herpesvirus 2) in the USA:molecular confirmation of isolates from diseased fish[J]. Journal of Aquatic Animal Health,2006,18(1):11-18.
[5] JUNG M H, RYU J W, NIKAPITIYA C, et al. Detection of herpesviral hematopoietic necrosis virus (cyprinid herpesvirus 2, CyHV-2) from goldfish, Carassius auratus (L. ) in Korea[J]. Fisheries and Aquatic Sciences,2022,25(7):403-408.
[6] ITO T, KURITA J, OZAKI A, et al. Growth of cyprinid herpesvirus 2 (CyHV-2) in cell culture and experimental infection of goldfish Carassius auratus[J]. Diseases of Aquatic Organisms,2013,105(3):193-202.
[7] WANG L, HE J, LIANG L, et al. Mass mortality caused by cyprinid herpesvirus 2 (CyHV-2) in prussian carp (Carassius gibelio) in China[J]. Bulletin of the European Association of Fish Pathologists, 2012,32(5):164-173.
[8] 吴霆,丁正峰,朱春艳,等.异育银鲫鳃出血病流行病学调查和研究[J].水产科学,2014,33(5):283-287.
[9] 李莉娟,罗杨志,刘学芹,等.金鱼鲤疱疹病毒Ⅱ型的分子诊断[J].华中农业大学学报,2013,32(1):92-96.
[10] 袁锐,刘训猛,陈静,等.江苏地区鲫鱼造血器官坏死病风险分析[J].安徽农业科学,2019,47(8):104-108.
[11] 赵哲,喻飞,郝凯,等.鱼类疱疹病毒研究进展概述[J].水产学报,2022,46(2):322-336.
[12] 罗丹,梁利国,谢骏,等.鲤疱疹病毒Ⅰ、Ⅱ、Ⅲ型研究进展[J].水生态学杂志,2014,35(3):94-100.
[13] GONCZOL E,MINAROVITS J, VALYI-NAGY T. Latency strategies of herpesviruses[M]. New York:Springer,2007.
[14] 唐睿哲.鲤疱疹病毒Ⅱ型极早期基因筛选及病毒感染对活性氧相关信号通路的影响[D].上海:上海海洋大学,2021:7-16.
[15] THANGARAJ R S, NITHIANANTHAM S R, DHARMARATNAM A, et al. Cyprinid herpesvirus-2 (CyHV-2):a comprehensive review[J]. Reviews in Aquaculture,2021,13(2):796-821.
[16] 孔善云,姜有声,许丹,等.一种基于Ⅱ型鲤疱疹病毒衣壳蛋白72的免疫学检测方法[J].中国水产科学,2016,23(2):328-335.
[17] 高娃,温虹,王浩,等.鲤疱疹病毒Ⅱ型主要免疫原性蛋白的鉴定[J].水产学报,2020,44(9):1441-1447.
[18] WALTZEK T B, KUROBE T, GOODWIN A E, et al. Development of a polymerase chain reaction assay to detect cyprinid herpesvirus 2 in goldfish[J]. Journal of Aquatic Animal Health,2009,21(1):60-67.
[19] 谢亚君,税典章,吴萍,等.基于不同PCR方法的Ⅱ型鲤疱疹病毒检测技术研究[J].基因组学与应用生物学,2019,38(3):1018-1025.
[20] GOODWIN A E, MERRY G E, SADLER J. Detection of the herpesviral hematopoietic necrosis disease agent (cyprinid herpesvirus 2) in moribund and healthy goldfish:validation of a quantitative PCR diagnostic method[J]. Diseases of Aquatic Organisms,2006,69(2/3):137-143.
[21] LIAN X H, SCOTT-THOMAS A, LEWIS J G, et al. Monoclonal antibodies and invasive aspergillosis:diagnostic and therapeutic perspectives[J]. International Journal of Molecular Sciences,2022,23(10):5563.
[22] WEI C, XU C N, SUN Y Y, et al. Investigation of the latency of cyprinid herpesvirus 2 in apparently healthy farmed gibel carp,Carassius auratus gibelio[J]. Aquaculture,2023,562:738854.
[23] LI Q,ZHAN W, XING J, et al. Production, characterisation and applicability of monoclonal antibodies to immunoglobulin of Japanese flounder (Paralichthys olivaceus)[J]. Fish & Shellfish Immunol,2007,23(5):982-990.
[24] 柴文君.鲤疱疹病毒Ⅱ潜伏感染的鉴定[D].武汉:华中农业大学,2019:4-8.
[25] WEI C, IIDA H, CHUAH Q, et al. Persistence of cyprinid herpesvirus 2 in asymptomatic goldfish Carassius auratus (L. ) that survived an experimental infection[J]. Journal of Fish Diseases,2019,42(6):913-921.
[26] WEI C, KAKAZU T, CHUAH Q Y, et al. Reactivation of cyprinid herpesvirus 2 (CyHV-2) in asymptomatic surviving goldfish Carassius auratus (L.) under immunosuppression[J]. Fish & Shellfish Immunology,2020,103:302-309.
[27] 李双,向涛,周海峰,等.鲤疱疹病毒Ⅱ型ORF72基因克隆、蛋白原核表达和抗体制备[J].江苏农业科学,2019,47(14):62-65.
[28] 曹欢,王静波,王姝,等.单克隆抗体技术在水生动物病害检测中的应用[J].北京农业,2015(2):80-83.
[29] 周勇,范玉顶,徐进,等.鲤疱疹病毒Ⅱ型ORF4基因的克隆、表达与免疫学检测方法[J].淡水渔业,2017,47(1):61-65.
[30] WU R H, ZHANG Q, LI Y. Development, characterization of monoclonal antibodies specific for the ORF25 membrane protein of cyprinid herpesvirus 2 and their applications in immunodiagnosis and neutralization of virus infection[J]. Aquaculture,2020,519:734904.
[31] 彭俊杰,张琪,贾路路,等.鲤疱疹病毒Ⅱ型(CyHV-2) ORF25蛋白的原核表达及单克隆抗体的制备[J].华中农业大学学报,2017,36(2):96-101.
[32] 高娃.鲤疱疹病毒Ⅱ型主要蛋白的研究与免疫检测试纸条的研制[D].上海:上海海洋大学,2021.
[33] MA J, JIANG N, LAPATRA S E, et al. Establishment of a novel and highly permissive cell line for the efficient replication of cyprinid herpesvirus 2 (CyHV-2)[J]. Veterinary Microbiology,2015,177(3/4):315-325.
[34] 许晨,周勇,史玉恒,等.鲫脑组织细胞系酵母双杂交cDNA文库的构建及初步应用[J].淡水渔业,2019,49(1):27-32.
[35] LU J F, XU D, LU L Q. A novel cell line established from caudal fin tissue of Carassius auratus gibelio is susceptible to cyprinid herpesvirus 2 infection with the induction of apoptosis[J]. Virus Research, 2018,258:19-27.
[36] SHIBATA T, NANJO A, SAITO M, et al. In vitro characteristics of cyprinid herpesvirus 2:effect of kidney extract supplementation on growth[J]. Diseases of Aquatic Organisms,2015,115(3):223-232.