Nanobody based immunoassay for detection of aquatic virus: Giant salamander iridovirus

文献类型： 外文期刊

第一作者： Xu, Zhihao

作者： Xu, Zhihao;He, Qiyi;Li, Wenkai;Pan, Junkang;Li, Dongyang;Xu, Zhihao;He, Qiyi;Li, Wenkai;Pan, Junkang;Li, Dongyang;Liu, Man;Liu, Wenzhi;El-Sheikh, El-Sayed A.;Hammock, Bruce D.;Hammock, Bruce D.

作者机构：

关键词： Aquaculture; Nanobody; ELISA; GSIV; polyHRP; Immunoassay

期刊名称：ANALYTICA CHIMICA ACTA （ 影响因子：6.0； 五年影响因子：5.7 ）

ISSN： 0003-2670

年卷期： 2025 年 1350 卷

页码：

收录情况： SCI

摘要： Aquatic viruses cause devastating diseases in aquaculture, severely limiting production and resulting in significant economic losses. The giant salamander iridovirus (GSIV), a member of the genus Ranavirus, is the only virus reported to infect giant salamanders, posing a severe threat to the farming industry. Currently, there are no effective strategies available for its control. In this context, a reliable diagnostic tool for the rapid detection of GSIV is crucial to mitigate its impact. This study developed a nanobody-based immunoassay for the rapid and reliable detection of GSIV. GSIV was cultured and used to immunize an alpaca, A phage library with an original diversity of 1.89 x 108 PFU was established, and six nanobodies were identified after three rounds of panning. Among them, HC-2 exhibited superior performance and was used to develop a highly sensitive ELISA method employing streptavidin-PolyHRP (SA-PolyHRP) as a signal amplification strategy. The assay achieved a detection limit of 3.3 x 105 PFU/mL and demonstrated high specificity without cross-reactivity. Practical application was validated in infected giant salamander samples, underscoring its diagnostic potential. This work provides a robust tool for GSIV diagnosis and showcases the potential of nanobodies in advancing aquaculture diagnostics.

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