Cuneyt Tamer, Hanne Nur Kuruçay, Seda Gözel, Bahadır Müftoglu, Elif Nur Kökluoğlu, Hamza Kadi, Fatih Dogan, Serdar Yildirim, Semra Gumusova, Zafer Yazici, Manfred Weidmann, Harun Albayrak
{"title":"重组酶聚合酶扩增驱动双侧流诊断试剂盒同时检测感染性胰腺坏死病毒和病毒性出血性败血症病毒:RPA和PCR方法的比较分析","authors":"Cuneyt Tamer, Hanne Nur Kuruçay, Seda Gözel, Bahadır Müftoglu, Elif Nur Kökluoğlu, Hamza Kadi, Fatih Dogan, Serdar Yildirim, Semra Gumusova, Zafer Yazici, Manfred Weidmann, Harun Albayrak","doi":"10.1111/jfd.70029","DOIUrl":null,"url":null,"abstract":"<p><p>Diagnosis time of infectious diseases in aquaculture is an important issue, and local and commercial enterprises generally do not prefer laboratory diagnosis of infectious diseases due to the long lead time and cost. This leads to the under-reporting of outbreaks in aquaculture farms and the recognition of infectious agents when they spread to many farm systems and cause significant economic loss. In this study, we aim to develop a fast and simple diagnostic test that can easily be applied under field conditions against Infectious pancreatic necrosis virus (IPNV) and Viral hemorrhagic septicemia virus (VHSV Ie), which are two crucial viral pathogens that cause significant problems in trout farming in Türkiye, both in hatcheries and farms. For this reason, we developed a lateral flow test strip for the duplex diagnosis of VHSV Ie and IPNV sp. based on Recombinase polymerase amplification (RPA), one of the molecular tests developed in recent years. We also compared Quantitative RT-PCR (q RT-PCR), Quantitative RT-RPA (rtRT-RPA) and lateral flow RT-RPA (lfRT-RPA) methods, which optimise both singleplex and duplex. As a result of this stage, the minimum detection limits of both rtRT-PCR, rtRT-RPA, and lfRT-RPA methods were determined. The lfRT-RPA test is almost as sensitive as rtRT-PCR and rtRT-RPA for the detection of duplex VHSV Ie, but for the detection of the IPNV sp., the lfRT-RPA test was found to be approximately 10-fold less sensitive than rtRT-PCR and rtRT-RPA. It is thought that the duplex lateral flow detection kit will allow timely diagnosis in the field for detecting infectious diseases of other aquaculture farms.</p>","PeriodicalId":15849,"journal":{"name":"Journal of fish diseases","volume":" ","pages":"e70029"},"PeriodicalIF":2.2000,"publicationDate":"2025-07-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Implementation of a Recombinase Polymerase Amplification-Driven Duplex Lateral Flow Diagnostic Kit for Concurrent Detecting Infectious Pancreatic Necrosis Virus and Viral Hemorrhagic Septicemia Virus: A Comparative Analysis of RPA and PCR Methods.\",\"authors\":\"Cuneyt Tamer, Hanne Nur Kuruçay, Seda Gözel, Bahadır Müftoglu, Elif Nur Kökluoğlu, Hamza Kadi, Fatih Dogan, Serdar Yildirim, Semra Gumusova, Zafer Yazici, Manfred Weidmann, Harun Albayrak\",\"doi\":\"10.1111/jfd.70029\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Diagnosis time of infectious diseases in aquaculture is an important issue, and local and commercial enterprises generally do not prefer laboratory diagnosis of infectious diseases due to the long lead time and cost. This leads to the under-reporting of outbreaks in aquaculture farms and the recognition of infectious agents when they spread to many farm systems and cause significant economic loss. In this study, we aim to develop a fast and simple diagnostic test that can easily be applied under field conditions against Infectious pancreatic necrosis virus (IPNV) and Viral hemorrhagic septicemia virus (VHSV Ie), which are two crucial viral pathogens that cause significant problems in trout farming in Türkiye, both in hatcheries and farms. For this reason, we developed a lateral flow test strip for the duplex diagnosis of VHSV Ie and IPNV sp. based on Recombinase polymerase amplification (RPA), one of the molecular tests developed in recent years. We also compared Quantitative RT-PCR (q RT-PCR), Quantitative RT-RPA (rtRT-RPA) and lateral flow RT-RPA (lfRT-RPA) methods, which optimise both singleplex and duplex. As a result of this stage, the minimum detection limits of both rtRT-PCR, rtRT-RPA, and lfRT-RPA methods were determined. The lfRT-RPA test is almost as sensitive as rtRT-PCR and rtRT-RPA for the detection of duplex VHSV Ie, but for the detection of the IPNV sp., the lfRT-RPA test was found to be approximately 10-fold less sensitive than rtRT-PCR and rtRT-RPA. It is thought that the duplex lateral flow detection kit will allow timely diagnosis in the field for detecting infectious diseases of other aquaculture farms.</p>\",\"PeriodicalId\":15849,\"journal\":{\"name\":\"Journal of fish diseases\",\"volume\":\" \",\"pages\":\"e70029\"},\"PeriodicalIF\":2.2000,\"publicationDate\":\"2025-07-29\",\"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.70029\",\"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.70029","RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"FISHERIES","Score":null,"Total":0}
Implementation of a Recombinase Polymerase Amplification-Driven Duplex Lateral Flow Diagnostic Kit for Concurrent Detecting Infectious Pancreatic Necrosis Virus and Viral Hemorrhagic Septicemia Virus: A Comparative Analysis of RPA and PCR Methods.
Diagnosis time of infectious diseases in aquaculture is an important issue, and local and commercial enterprises generally do not prefer laboratory diagnosis of infectious diseases due to the long lead time and cost. This leads to the under-reporting of outbreaks in aquaculture farms and the recognition of infectious agents when they spread to many farm systems and cause significant economic loss. In this study, we aim to develop a fast and simple diagnostic test that can easily be applied under field conditions against Infectious pancreatic necrosis virus (IPNV) and Viral hemorrhagic septicemia virus (VHSV Ie), which are two crucial viral pathogens that cause significant problems in trout farming in Türkiye, both in hatcheries and farms. For this reason, we developed a lateral flow test strip for the duplex diagnosis of VHSV Ie and IPNV sp. based on Recombinase polymerase amplification (RPA), one of the molecular tests developed in recent years. We also compared Quantitative RT-PCR (q RT-PCR), Quantitative RT-RPA (rtRT-RPA) and lateral flow RT-RPA (lfRT-RPA) methods, which optimise both singleplex and duplex. As a result of this stage, the minimum detection limits of both rtRT-PCR, rtRT-RPA, and lfRT-RPA methods were determined. The lfRT-RPA test is almost as sensitive as rtRT-PCR and rtRT-RPA for the detection of duplex VHSV Ie, but for the detection of the IPNV sp., the lfRT-RPA test was found to be approximately 10-fold less sensitive than rtRT-PCR and rtRT-RPA. It is thought that the duplex lateral flow detection kit will allow timely diagnosis in the field for detecting infectious diseases of other aquaculture farms.
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