| 摘要: |
| 石斑鱼虹彩病毒 (Singapore grouper iridovirus，SGIV)是海水养殖中危害严重的病原体，即时检测技术的研发对于及时精准防控病原意义重大。然而，传统检测方法存在操作复杂、耗时长、依赖精密设备等局限，难以满足养殖现场快速诊断需求。因此亟需建立一种快速、灵敏、简便的检测方法，以满足水产养殖现场对病毒早期即时诊断的需求。本研究根据实验室前期获得的可特异性识别SGIV感染的核酸适体LYGV1，将其进行一定的截短及修饰，以构建一种快速的检测方法。通过光学显微镜观察、细胞活力测定、荧光定量PCR以及流式细胞仪等方法进行验证，结果发现，Cy5-LYGV1c在浓度为1000 nM，4~28 ℃温度范围内短至1 min内既可以检测到5×103 个/mL SGIV感染的细胞。活体实验显示，Cy5-LYGV1c能特异性识别SGIV感染的样本，其检测结果与RT-qPCR结果一致，证实Cy5-LYGV1c的可靠性。且Cy5-LYGV1c的整个检测流程仅需要2 h即可完成，大大缩短了检测时间。综上所述，本研究建立了一种耗时短、特异性强、灵敏度高、操作简便的SGIV检测技术，说明该技术有望进一步开发为现场快速检测试剂盒，并为构建“检测—治疗”一体化防控平台提供技术基础。 |
| 关键词: 石斑鱼虹彩病毒 海水养殖 核酸适体 检测技术 |
| DOI： |
| 投稿时间：2026-03-09修订日期：2026-04-27 |
| 基金项目:广西自然科学(AD23026331,2023GXNSFAA026325)；广西科技计划项目(桂科发[2024]102-2)；来宾市科学研究与技术开发计划项目(来科产241519)；广东海洋大学研究生教育创新计划资助项目(202422) |
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| Rapid Detection Technology for Grouper Iris Virus Based on the Specific Recognition of the Nucleic Acid Aptamer LYGV1c |
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gaoyu1,2, yuqing3,4, xujiwei5, susining6, caijia7, lipengfei7,2,4, huangjing2,4, liumingzhu8,2
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| (1. Guangdong Provincial Key Laboratory of Aquatic Animal Disease Control and Healthy Culture, Fisheries College, Guangdong Ocean University, Zhanjiang, P.R. China;2. Guangxi Academy of Marine Sciences, Guangxi Key Laboratory of Aquatic Biotechnology and Modern Ecological Aquaculture, Guangxi Engineering Research Center for Fishery Major Diseases Control and Efficient Healthy Breeding Industrial Technology (GERCFT), Guangxi Academy of Sciences, Nanning, P.R. China;3.. Guangxi Academy of Marine Sciences, Guangxi Key Laboratory of Aquatic Biotechnology and Modern Ecological Aquaculture, Guangxi Engineering Research Center for Fishery Major Diseases Control and Efficient Healthy Breeding Industrial Technology (GERCFT), Guangxi Academy of Sciences, Nanning, P.R. China;4.5. Guangxi Doctoral Innovation Station, Guangxi Fuqun Seawater Seedling Propagation Co. LTD, Fangchenggang, P.R. China;5.Guangxi Doctoral Innovation Station, Guangxi Ruona Biotechnology Co. LTD, Fangchenggang, P.R. China;6.Guangxi Doctoral Innovation Station, Guangxi Fuqun Seawater Seedling Propagation Co. LTD, Fangchenggang, P.R. China;7.Guangdong Provincial Key Laboratory of Aquatic Animal Disease Control and Healthy Culture, Fisheries College, Guangdong Ocean University, Zhanjiang, P.R. China;8.1.Guangdong Provincial Key Laboratory of Aquatic Animal Disease Control and Healthy Culture, Fisheries College, Guangdong Ocean University, Zhanjiang, P.R. China) |
| Abstract: |
| Singapore grouper iridovirus (SGIV) is a highly pathogenic agent in mariculture, and the development of rapid detection technologies is crucial for timely and accurate disease control. However, traditional detection methods suffer from limitations such as complex procedures, time-consuming processes, and reliance on sophisticated equipment, making it difficult to meet the demand for rapid on-site diagnosis in aquaculture settings. Therefore, there is an urgent need to establish a rapid, sensitive, and simple detection method to meet the demand for early, on-site diagnosis of the virus in aquaculture. In this study, based on the nucleic acid aptamer LYGV1—previously obtained in the laboratory and capable of specifically recognizing SGIV infection—we truncated and modified it to construct a rapid detection method. Validation was conducted using optical microscopy, cell viability assays, quantitative real-time PCR (qPCR), and flow cytometry. The results showed that Cy5-LYGV1c could detect as few as 5 × 103 SGIV-infected cells per mL within 1 minute at a concentration of 1000 nM and temperatures ranging from 4 to 28 °C. In vivo experiments demonstrated that Cy5-LYGV1c specifically recognizes SGIV-infected samples, and its detection results were consistent with RT-qPCR findings, confirming the reliability of Cy5-LYGV1c. Furthermore, the entire detection process for Cy5-LYGV1c can be completed in just 2 hours, significantly reducing the detection time. In summary, this study established a time-efficient, highly specific, and sensitive SGIV detection technology that is easy to operate. This suggests that the technology holds promise for further development into a rapid point-of-care diagnostic kit and provides a technical foundation for building an integrated “detection-treatment” prevention and control platform prevention and control platform. |
| Key words: Singapore grouper iridovirus Mariculture Aptamer Detection technology |
