{"title":"RPA联合LFS快速检测十足虫彩虹病毒1型（DIV1）","authors":"Quanling Mu, Cunbao Ding, Ying Xie, Xi Zhen, Xianrui Wang, Tongaobo Qi, Jiaming Zhang, Yakun Yu","doi":"10.1111/jfd.70052","DOIUrl":null,"url":null,"abstract":"<p><p>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<sub>AHPND</sub>. In addition, RPA-LFS has high detection sensitivity, with a lower detection limit of 1.12 × 10<sup>1</sup> 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.</p>","PeriodicalId":15849,"journal":{"name":"Journal of fish diseases","volume":" ","pages":"e70052"},"PeriodicalIF":2.2000,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Rapid Visual Detection of Decapod Iridescent Virus 1 (DIV1) by RPA Combined With LFS.\",\"authors\":\"Quanling Mu, Cunbao Ding, Ying Xie, Xi Zhen, Xianrui Wang, Tongaobo Qi, Jiaming Zhang, Yakun Yu\",\"doi\":\"10.1111/jfd.70052\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>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<sub>AHPND</sub>. In addition, RPA-LFS has high detection sensitivity, with a lower detection limit of 1.12 × 10<sup>1</sup> 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.</p>\",\"PeriodicalId\":15849,\"journal\":{\"name\":\"Journal of fish diseases\",\"volume\":\" \",\"pages\":\"e70052\"},\"PeriodicalIF\":2.2000,\"publicationDate\":\"2025-09-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of fish diseases\",\"FirstCategoryId\":\"97\",\"ListUrlMain\":\"https://doi.org/10.1111/jfd.70052\",\"RegionNum\":3,\"RegionCategory\":\"农林科学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"FISHERIES\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of fish diseases","FirstCategoryId":"97","ListUrlMain":"https://doi.org/10.1111/jfd.70052","RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"FISHERIES","Score":null,"Total":0}
Rapid Visual Detection of Decapod Iridescent Virus 1 (DIV1) by RPA Combined With LFS.
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 VpAHPND. 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.
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
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