Citation: | LI Shuo, CHEN Jingni, ZHAO Lining, HUANG Chunping, HUANG Jinlu, WANG Guiping, ZHONG Ying. Preparation of anti-largemouth bass ranavirus egg yolk antibody and establishment of indirect ELISA method[J]. South China Fisheries Science, 2024, 20(2): 129-139. DOI: 10.12131/20230148 |
Largemouth bass ranavirus (LMBV) is a major pathogenic agent in largemouth bass culture in China, which mainly causes viral canker disease and restricts the healthy development of largemouth bass culture. In order to explore the potential role of egg yolk antibody in the prevention and control of LMBV, we immunized the LMBV inactivated vaccine to laying hens and prepared the egg yolk immunoglobulin against LMBV (Anti-LMBV IgY). Besides, we established an indirect enzyme linked immuno sorbent assay (ELISA) method for detecting the titer of anti-LMBV IgY by screening trapping concentration, encapsulation condition and sealing condition. Then we evaluated the titer of anti-LMBV IgY at different time points post LMBV inactivated vaccine immunization by using the established method. The results show that 1‰ β-propanolactone could inactivate LMBV at 4 ℃ for 72 h completely. For indirect ELISA, being coated with 105 TCID50 inactivated virus, incubated at 37 ℃ for 2 h, and blocked with 5% bovine serum albumin at 37 ℃ for 2 h could reduce the background values of negative control effectively. In addition, no cross reaction had been detected between inactivate LMBV with other egg yolk extract or anti-LMBV IgY with cells, indicating that the indirect ELISA method had high specificity. Finally, we detected specific anti-LMBV IgY titer at 48 d post LMBV inactivated vaccine immunization; the titer increased to a peak of 1:12800 after 58 d post immunization, and last up to 128 d. To sum up, the indirect ELISA method in this study can real-time monitor the titer level of anti-LMBV IgY, determine the efficient immunization program and the duration of high-immunity egg collection, facilitate the development and application of LMBV egg yolk antibody products, and provide a potential solution for the prevention and treatment of LMBV.
[1] |
李江涛, 杨凯宇, 邱晓桐, 等. 大口黑鲈佛山和台湾群体自交与杂交子代的生长和形态差异分析[J]. 南方水产科学, 2021, 17(5): 1-9.
|
[2] |
农业农村部渔业渔政管理局, 全国水产技术推广总站, 中国水产学会. 2023中国渔业统计年鉴[M]. 北京: 中国农业出版社, 2023: 33.
|
[3] |
LIU X D, ZHANG Y B, ZHANG Z L, et al. Isolation, identification and the pathogenicity characterization of a Santee-Cooper ranavirus and its activation on immune responses in juvenile largemouth bass (Micropterus salmoides)[J]. Fish Shellfish Immunol, 2023, 135: 108641. doi: 10.1016/j.fsi.2023.108641
|
[4] |
杨超, 董浚键, 刘志刚, 等. 大口黑鲈源维氏气单胞菌的分离鉴定[J]. 南方水产科学, 2021, 17(3): 54-61. doi: 10.12131/20200230
|
[5] |
BAI J J, LI J J. Development of largemouth bass (Micropterus salmoides) culture[M]. New Jersey: John Wiley & Sons, Ltd. , 2018: 421-429.
|
[6] |
ZHAO R X, GENG Y, QIN Z Y, et al. A new ranavirus of the Santee-Cooper group invades largemouth bass (Micropterus salmoides) culture in southwest China[J]. Aquaculture, 2020, 526(1): 735363.
|
[7] |
邓国成, 谢骏, 李胜杰, 等. 大口黑鲈病毒性溃疡病的病原分离和鉴定初步研究[J]. 水产学报, 2009, 33(5): 871-877.
|
[8] |
马冬梅, 邓国成, 白俊杰, 等. 大口黑鲈肝脾肿大病病原研究[J]. 中国水产科学, 2011, 18(3): 654-660.
|
[9] |
马冬梅, 白俊杰, 邓国成, 等. 大口黑鲈溃疡综合征病毒MCP基因序列分析及PCR快速检测方法的建立[J]. 中国水产科学, 2010, 17(6): 1149-1156.
|
[10] |
YI, W Y, ZHANG X, ZENG K, et al. Construction of a DNA vaccine and its protective effect on largemouth bass (Micropterus salmoides) challenged with largemouth bass virus (LMBV)[J]. Fish Shellfish Immunol, 2020, 106: 103-109. doi: 10.1016/j.fsi.2020.06.062
|
[11] |
JIA Y J, GUO Z R, MA R, et al. Protective immunity of largemouth bass immunized with immersed DNA vaccine against largemouth bass ulcerative syndrome virus[J]. Fish Shellfish Immunol, 2020, 107(Pt A): 269-276.
|
[12] |
JIA Y J, GUO Z R, MA R, et al. Immune efficacy of carbon nanotubes recombinant subunit vaccine against largemouth bass ulcerative syndrome virus[J]. Fish Shellfish Immunol, 2020, 100: 317-323. doi: 10.1016/j.fsi.2020.03.025
|
[13] |
YAO J Y, ZHANG C S, YUAN X M, et al. Oral vaccination with recombinant Pichia pastoris expressing iridovirus major capsid protein elicits protective immunity in largemouth bass (Micropterus salmoides)[J]. Front Immunol, 2022, 13: 852300. doi: 10.3389/fimmu.2022.852300
|
[14] |
ZHANG M J, CHEN X Y, XUE M Y, et al. Oral vaccination of largemouth bass (Micropterus salmoides) against largemouth bass ranavirus (LMBV) using yeast surface display technology[J]. Animals (Basel), 2023, 13(7): 1183.
|
[15] |
徐永平, 徐乐, 李纪彬, 等. 卵黄抗体饲料添加剂在畜牧及水产养殖病害防控中的研究进展[J]. 饲料工业, 2021, 42(18): 1-11.
|
[16] |
XU Y P, LI X Y, JIN L J, et al. Application of chicken egg yolk immunoglobulins in the control of terrestrial and aquatic animal diseases: a review[J]. Biotechnol Adv, 2011, 29(6): 860-868. doi: 10.1016/j.biotechadv.2011.07.003
|
[17] |
MINE Y, KOVACS-NOLAN J. Chicken egg yolk antibodies as therapeutics in enteric infectious disease: a review[J]. J Med Food, 2002, 5(3): 159-169. doi: 10.1089/10966200260398198
|
[18] |
翟玥, 曲笑锋, 庞博, 等. 副溶血性弧菌高免卵黄抗体的制备和不同提纯方法效果的比较[J]. 吉林大学学报(医学版), 2017, 43(2): 441-445. doi: 10.13481/j.1671-587x.20170245
|
[19] |
杨帆, 张连峰. 卵黄抗体的分离提取和纯化方法[J]. 中国比较医学杂志, 2007, 17(11): 688-689, 693.
|
[20] |
甄宇红, 徐永平, 夏艳秋, 等. 卵黄抗体分离、提取和纯化方法研究进展[J]. 中国饲料, 2003(18): 8-10. doi: 10.3969/j.issn.1004-3314.2003.18.004
|
[21] |
XU L, CHE J, XU Y P, et al. Oral administration of microencapsulated egg yolk immunoglobulin (IgY) in turbot (Scophthalmus maximus) to combat against Edwardsiella tarda 2CDM001 infections[J]. Fish Shellfish Immunol, 2020, 106: 609-620. doi: 10.1016/j.fsi.2020.08.024
|
[22] |
DIAS da SILVA W, TAMBOURGI D V. IgY: a promising antibody for use in immunodiagnostic and in immunotherapy[J]. Vet Immunol Immunopathol, 2010, 135(3/4): 173-180.
|
[23] |
张绍兰. 鸡卵黄抗体研究进展[J]. 西南军医, 2008(3): 120-121. doi: 10.3969/j.issn.1672-7193.2008.03.078
|
[24] |
YAKHKESHI S, WU R, CHELLIAPPAN B, et al. Trends in industrialization and commercialization of IgY technology[J]. Front Immunol, 2022, 13: 991931. doi: 10.3389/fimmu.2022.991931
|
[25] |
LU Y N, LIU J J, JIN L J, et al. Passive immunization of crayfish (Procambius clarkiaii) with chicken egg yolk immunoglobulin (IgY) against white spot syndrome virus (WSSV)[J]. Appl Biochem Biotechnol, 2009, 159(3): 750-758. doi: 10.1007/s12010-009-8555-6
|
[26] |
LU Y N, LIU J J, JIN L J, et al. Passive protection of shrimp against white spot syndrome virus (WSSV) using specific antibody from egg yolk of chickens immunized with inactivated virus or a WSSV-DNA vaccine[J]. Fish Shellfish Immunol, 2008, 25(5): 604-610. doi: 10.1016/j.fsi.2008.08.010
|
[27] |
YI L Z, QIN Z D, LIN H Z, et al. Features of chicken egg yolk immunoglobulin (IgY) against the infection of red-spotted grouper nervous necrosis virus[J]. Fish Shellfish Immunol, 2018, 80: 534-539. doi: 10.1016/j.fsi.2018.06.024
|
[28] |
寇海燕, 郭培红, 田丹阳, 等. 抗鲤疱疹Ⅱ型病毒卵黄抗体制备及其功能鉴定[J]. 淡水渔业, 2021, 51(1): 65-71. doi: 10.3969/j.issn.1000-6907.2021.01.008
|
[29] |
袁雪梅, 吕孙建, 施伟达, 等. 大口黑鲈弹状病毒的分离培养及其卵黄抗体的制备[J]. 渔业科学进展, 2020, 41(3): 151-157. doi: 10.19663/j.issn2095-9869.20190704001
|
[30] |
HU B C, YANG X D, GUO E P, et al. The preparation and antibacterial effect of egg yolk immunoglobulin (IgY) against the outer membrane proteins of Vibrio parahaemolyticus[J]. J Sci Food Agric, 2019, 99(5): 2565-2571. doi: 10.1002/jsfa.9470
|
[31] |
NAKAMURA R, PEDROSA-GERASMIO I R, ALENTON R R R, et al. Anti-PirA-like toxin immunoglobulin (IgY) in feeds passively immunizes shrimp against acute hepatopancreatic necrosis disease[J]. J Fish Dis, 2019, 42(8): 1125-1132. doi: 10.1111/jfd.13024
|
[32] |
GRIZZLE J M, ALTINOK I, FRASER W A, et al. First isolation of largemouth bass virus[J]. Dis Aquat Organ, 2002, 50(3): 233-235.
|
[33] |
张小莺, 郑礼, 等. 免疫鸡产生IgY抗体的技术[J]. 中国药理学通报, 2004, 20(10): 1102-1106. doi: 10.3321/j.issn:1001-1978.2004.10.006
|
[34] |
郝丽芳, 邱宁, 马美湖, 等. 鸡蛋黄中蛋白质研究进展[J]. 中国家禽, 2012, 34(21): 45-50. doi: 10.3969/j.issn.1004-6364.2012.21.012
|
[35] |
王洪新, 刘学贤, 穆海波, 等. 鸡新城疫卵黄抗体IgY的分离提纯研究[J]. 中国家禽, 2003, 25(19): 14-17. doi: 10.3969/j.issn.1004-6364.2003.19.004
|
[36] |
VEERASAMI M, SINGANALLUR N B, THIRUMENI N, et al. Serotyping of foot-and-mouth disease virus by antigen capture-ELISA using monoclonal antibodies and chicken IgY[J]. New Microbiol, 2008, 31(4): 549-554.
|
[37] |
王云芸. 抗诺如病毒鸡卵黄抗体的制备及其初步应用研究[D]. 广州: 南方医科大学, 2013: 34-54.
|
[38] |
张婧兰. 犬瘟热病毒和犬细小病毒高免卵黄抗体的制备及其壳聚糖微球的研究[D]. 南京: 南京农业大学, 2009: 30-36.
|
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