Mass mortality of mandarin fish(Siniperca chuatsi)due to infectious spleen and kidney necrosis virus(ISKNV)infection occurs frequently.Since there are no effective drug treatments available,prevention relies heavily o...Mass mortality of mandarin fish(Siniperca chuatsi)due to infectious spleen and kidney necrosis virus(ISKNV)infection occurs frequently.Since there are no effective drug treatments available,prevention relies heavily on detection.Effective and rapid on-site detection methods are needed for early diagnosis of ISKNV.In this study,a rapid and simple colloidal gold test strip method,specific for the antibody against major capsid protein(MCP),was developed and systematically evaluated for the detection of ISKNV.The limit of detection of the test strip is a 1꞉100 dilution of a positive standard serum and the antibody level in the fish could be estimated from the depth of color of the test line.The strips were tested against serum samples of cyprinid herpesvirus-2(CyHV-2),grass carp reovirus(GCRV),largemouth bass ranavirus(LMBV),large yellow croaker iridovirus(LYCIV),and spring viremia of carp virus(SVCV),yielding no cross-reactivity.In addition,10 mandarin fish artificially infected with ISKNV were tested using the current industry standard PCR method(SC/T 7211-2011)on their splenorenal tissues.The results from the test strips showed a high degree of concordance with PCR testing,achieving a Kappa value of 0.737.All the results indicated that the colloidal gold test strips prepared in this study could be used as a simple,rapid,and highly sensitive and specific method for ISKNV diagnosis.展开更多
传染性脾肾坏死病毒(ISKNV)是鳜鱼重大疫病病原,该病毒引发的鳜鱼传染性脾肾坏死病(Infection spleen and kidney necrosis)能导致养殖生产巨大经济损失。为满足水产疫病预防与诊断需求,该文将一种集DNA快速提取与PCR快速扩增为一体的...传染性脾肾坏死病毒(ISKNV)是鳜鱼重大疫病病原,该病毒引发的鳜鱼传染性脾肾坏死病(Infection spleen and kidney necrosis)能导致养殖生产巨大经济损失。为满足水产疫病预防与诊断需求,该文将一种集DNA快速提取与PCR快速扩增为一体的分子生物学检测方法与试剂盒提取DNA法和传染性脾肾坏死病毒检测方法(SC/T7211-2011)中PCR方法进行比较,旨在为基层水产一线水生动物疫病预防与控制提供一种快速检测方法。展开更多
A gene library of the infectious spleen and kidney necrosis virus(ISKNV) from mandarinfish,Siniperca chuatsi(Basilewsky) genome was established ISKNV DNA was cleaved with EcoRI,BamHI,HindⅢ,KpnI and PstI and the resul...A gene library of the infectious spleen and kidney necrosis virus(ISKNV) from mandarinfish,Siniperca chuatsi(Basilewsky) genome was established ISKNV DNA was cleaved with EcoRI,BamHI,HindⅢ,KpnI and PstI and the resulting fragments were inserted into the corresponding sites of the pBluescript Ⅱ KS plasmid vector using T4 DNA ligase Bacterial colonies harboring recombinant plasmids were selected All cloned fragments were individually identified by digestion of the recombinant plasmid DNA with different restriction enzymes and screened by hybridization of recombinant plasmid DNA to viral DNA Using these recombinant plasmids,the physical maps of the genome were constructed for BamHI,HindⅢ and KpnI restriction展开更多
基金Supported by the Earmarked Fund for China Agricultural Research System(No.CARS-45-16)the Key Research and Development Plan of Jiangsu Province(No.BE2021369)。
文摘Mass mortality of mandarin fish(Siniperca chuatsi)due to infectious spleen and kidney necrosis virus(ISKNV)infection occurs frequently.Since there are no effective drug treatments available,prevention relies heavily on detection.Effective and rapid on-site detection methods are needed for early diagnosis of ISKNV.In this study,a rapid and simple colloidal gold test strip method,specific for the antibody against major capsid protein(MCP),was developed and systematically evaluated for the detection of ISKNV.The limit of detection of the test strip is a 1꞉100 dilution of a positive standard serum and the antibody level in the fish could be estimated from the depth of color of the test line.The strips were tested against serum samples of cyprinid herpesvirus-2(CyHV-2),grass carp reovirus(GCRV),largemouth bass ranavirus(LMBV),large yellow croaker iridovirus(LYCIV),and spring viremia of carp virus(SVCV),yielding no cross-reactivity.In addition,10 mandarin fish artificially infected with ISKNV were tested using the current industry standard PCR method(SC/T 7211-2011)on their splenorenal tissues.The results from the test strips showed a high degree of concordance with PCR testing,achieving a Kappa value of 0.737.All the results indicated that the colloidal gold test strips prepared in this study could be used as a simple,rapid,and highly sensitive and specific method for ISKNV diagnosis.
文摘传染性脾肾坏死病毒(ISKNV)是鳜鱼重大疫病病原,该病毒引发的鳜鱼传染性脾肾坏死病(Infection spleen and kidney necrosis)能导致养殖生产巨大经济损失。为满足水产疫病预防与诊断需求,该文将一种集DNA快速提取与PCR快速扩增为一体的分子生物学检测方法与试剂盒提取DNA法和传染性脾肾坏死病毒检测方法(SC/T7211-2011)中PCR方法进行比较,旨在为基层水产一线水生动物疫病预防与控制提供一种快速检测方法。
文摘A gene library of the infectious spleen and kidney necrosis virus(ISKNV) from mandarinfish,Siniperca chuatsi(Basilewsky) genome was established ISKNV DNA was cleaved with EcoRI,BamHI,HindⅢ,KpnI and PstI and the resulting fragments were inserted into the corresponding sites of the pBluescript Ⅱ KS plasmid vector using T4 DNA ligase Bacterial colonies harboring recombinant plasmids were selected All cloned fragments were individually identified by digestion of the recombinant plasmid DNA with different restriction enzymes and screened by hybridization of recombinant plasmid DNA to viral DNA Using these recombinant plasmids,the physical maps of the genome were constructed for BamHI,HindⅢ and KpnI restriction
