Rapid Visual Detection of Decapod Iridescent Virus 1 (DIV1) by RPA Combined With LFS.

IF 2.2 3区农林科学 Q2 FISHERIES

Journal of fish diseases Pub Date : 2025-09-01 DOI:10.1111/jfd.70052

Quanling Mu, Cunbao Ding, Ying Xie, Xi Zhen, Xianrui Wang, Tongaobo Qi, Jiaming Zhang, Yakun Yu

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引用次数: 0

Abstract

Decapod iridescent virus 1 (DIV1) causes severe disease outbreaks in shrimp and crab culture areas. A simple, rapid, and visual DIV1 assay is important for the control of viral diseases. This study presented a novel DIV1 detection method that combines recombinase polymerase amplification (RPA) and lateral flow strip (LFS). After selecting primers and probes, we optimised the concentration of the reverse primers, reaction time, as well as reaction temperature of RPA-LFS detection. RPA can amplify the target gene within 18 min at a constant temperature of 38°C, and LFS can observe the amplification results within 3 min. Importantly, there is no cross-reactivity with other infectable shrimp viruses and pathogens, such as WSSV, IHHNV, TSV, EHP, CMNV, YHV, MrNV samples, as well as Vp_AHPND. In addition, RPA-LFS has high detection sensitivity, with a lower detection limit of 1.12 × 10¹ copies/μL. Using 110 field samples, the results of qPCR recommended by WOAH (OIE) and RPA-LFS were identical, indicating that RPA-LFS is as reliable as qPCR. The RPA-LFS assay is a valuable tool for the rapid and accurate detection of DIV1.

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RPA联合LFS快速检测十足虫彩虹病毒1型（DIV1）

十足类虹彩病毒1 （DIV1）在虾蟹养殖区引起严重的疾病暴发。一种简单、快速、直观的DIV1检测方法对病毒性疾病的控制具有重要意义。本研究提出了一种结合重组酶聚合酶扩增（RPA）和横向流动条带（LFS）检测DIV1的新方法。在选择引物和探针后，对RPA-LFS检测的反引物浓度、反应时间和反应温度进行优化。在38℃恒温条件下，RPA可在18 min内扩增目的基因，LFS可在3 min内观察扩增结果。重要的是，与WSSV、IHHNV、TSV、EHP、CMNV、YHV、MrNV样品以及VpAHPND等其他传染性虾病毒和病原体无交叉反应性。RPA-LFS具有较高的检测灵敏度，检测限为1.12 × 101拷贝/μL。在110份野外样品中，WOAH （OIE）推荐的qPCR结果与RPA-LFS结果一致，表明RPA-LFS与qPCR一样可靠。RPA-LFS是快速、准确检测DIV1的一种有价值的工具。

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来源期刊

Journal of fish diseases 农林科学-海洋与淡水生物学

CiteScore

4.60

自引率

12.00%

发文量

170

审稿时长

6 months

期刊介绍： Journal of Fish Diseases enjoys an international reputation as the medium for the exchange of information on original research into all aspects of disease in both wild and cultured fish and shellfish. Areas of interest regularly covered by the journal include: -host-pathogen relationships- studies of fish pathogens- pathophysiology- diagnostic methods- therapy- epidemiology- descriptions of new diseases