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基于核酸适体LYGV1c的酶联吸附法检测石斑鱼虹彩病毒感染

刘明珠陈嘉余庆李鹏飞黄静程远韦云依牟容丽黄琳竺利波陆兰天柯珂

广东海洋大学学报2024，Vol.44Issue(3)：1-8,8.

广东海洋大学学报2024，Vol.44Issue(3)：1-8,8.DOI:10.3969/j.issn.1673-9159.2024.03.001

基于核酸适体LYGV1c的酶联吸附法检测石斑鱼虹彩病毒感染

Detection of Grouper Iridovirus by Enzyme-linked Adsorption Method Based on Aptamer LYGV1c

刘明珠 ¹陈嘉 ¹余庆 ¹李鹏飞 ²黄静 ¹程远 ¹韦云依 ³牟容丽 ¹黄琳 ¹竺利波 ⁴陆兰天 ⁴柯珂⁴

作者信息

1. 广西水产生物技术与现代生态养殖重点实验室,广西渔业重大疫病防控与高效健康养殖产业技术工程研究中心,广西科学院,广西南宁 530007||中国-东盟现代渔业产业技术转移示范中心,广西科学院北部湾海洋产业研究院,广西南宁 530007
2. 广西水产生物技术与现代生态养殖重点实验室,广西渔业重大疫病防控与高效健康养殖产业技术工程研究中心,广西科学院,广西南宁 530007||南宁学院食品与质量工程学院,广西南宁 530200||中国-东盟现代渔业产业技术转移示范中心,广西科学院北部湾海洋产业研究院,广西南宁 530007
3. 南宁学院食品与质量工程学院,广西南宁 530200
4. 广西水产生物技术与现代生态养殖重点实验室,广西渔业重大疫病防控与高效健康养殖产业技术工程研究中心,广西科学院,广西南宁 530007
折叠

摘要

Abstract

[Objective]To develop the aptamer LYGV1c-based enzyme-linked apta-sorbent assay(LYGV1c-ELASA)for accurate and rapid detection of Singapore grouper iridovirus Guangxi strain(SGIV-Gx),and to provide theoretical support for the development of rapid detection system technology in aquaculture.[Method]An aptamer enzyme-linked adsorption method was developed based on biotin-labeled LYGV1c.The specificity,sensitivity and stability of bio-LYGV1c were analyzed in SGIV-Gx infection.Hybrid Grouper(Epinephelus fuscoguttatus♀×Epinephelus lanceolatus♂)were infected by SGIV-Gx,and the relative expression of MCP of SGIV-Gx was determined by fluorescence quantitative PCP(qPCR),the results of qPCR were compared with that of LYGV1c-ELASA to verify the reliability of ELASA.[Result]LYGV1c-ELASA could detect SGIV infection with high specificity,and the working concentration was 500 nmol/L,incubation was 20 min,the suitable binding temperature was 4‒28℃,and the detection limit was 5×103 cells/mL.The results of infection test showed that the OD450 value of SGIV-Gx increased with the increase of virus concentration in vivo.The qPCR result was the same as LYGV1c-ELASA detection results in vivo.[Conclusion]LYGV1c-ELASA technology can be used for the detection of SGIV-Gx in vitro and in vivo.LYGV1c-ELASA is a technique for rapid diagnosis and real-time monitor of SGIV-Gx infection.

关键词

石斑鱼虹彩病毒/检测/核酸适体/酶联吸附法

Key words

grouper iridovirus/detection/aptamer/enzyme-linked apta-sorbent assay(ELASA)

分类

农业科技

引用本文复制引用

刘明珠,陈嘉,余庆,李鹏飞,黄静,程远,韦云依,牟容丽,黄琳,竺利波,陆兰天,柯珂..基于核酸适体LYGV1c的酶联吸附法检测石斑鱼虹彩病毒感染[J].广东海洋大学学报,2024,44(3):1-8,8.

基金项目

国家自然科学基金(32373175) （32373175）

广西自然科学基金(2023JJG130007,2022GXNSFBA035521,2022JJA130074) （2023JJG130007,2022GXNSFBA035521,2022JJA130074）

国家重点研发计划(2022YFD2401200) （2022YFD2401200）

国家现代农业产业技术体系广西创新团队项目(nycytxgxcxtd-2021-08-02). （nycytxgxcxtd-2021-08-02）

广东海洋大学学报

OA北大核心CHSSCDCSTPCD

ISSN：1673-9159

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