鲤春病毒血症(spring virernia of carp,SVC)是由鲤春病毒血症病毒(carp spring virernia virus, CSVV)引起的多种鲤科鱼类高死亡率的病毒病,一般发生在春季,暴发后世界渔业经济损失严重。SVC是世界动物卫生组织(Office International D...鲤春病毒血症(spring virernia of carp,SVC)是由鲤春病毒血症病毒(carp spring virernia virus, CSVV)引起的多种鲤科鱼类高死亡率的病毒病,一般发生在春季,暴发后世界渔业经济损失严重。SVC是世界动物卫生组织(Office International Des Epizooties,OIE)必报的重要疫病,也是我国2008年最新颁布的动物疫病病种名录中的一类动物疫病。本文阐述了CSVV的病原生物学特性、流行病学、病理学,介绍国内外CSVV的检测技术手段、防治方法及疫苗的研发等,为CSVV的防治提供参考。展开更多
糖蛋白(Glycoprotein,G)作为鲤春病毒血症病毒(Spring Virernia of Carp Virus,SVCV)主要的抗原蛋白,已成为现阶段SVCV病毒检测、抗体制备以及疫苗研制的热点。为了对其进行酵母表面展示,研究以SVCVshlj1分离株基因组为模板,通过RT-PCR...糖蛋白(Glycoprotein,G)作为鲤春病毒血症病毒(Spring Virernia of Carp Virus,SVCV)主要的抗原蛋白,已成为现阶段SVCV病毒检测、抗体制备以及疫苗研制的热点。为了对其进行酵母表面展示,研究以SVCVshlj1分离株基因组为模板,通过RT-PCR技术,体外扩增获得SVCV表面糖蛋白的基因开放阅读框(1530 bp)片段,将其克隆至酵母表面展示载体pYD1,构建重组质粒pYD1-G。利用电转化方法将重组质粒pYD1-G导入酿酒酵母EBY100感受态细胞,经YNB选择培养基筛选和菌液PCR的鉴定,挑选出阳性转化子(命名为EBY100-pYD1-G),对其进行2%半乳糖诱导。利用细胞免疫荧光和流式细胞仪检测G蛋白的酵母表面展示情况。细胞免疫荧光结果显示,诱导后的酵母细胞EBY100-pYD1-G能产生特异性红色荧光,且随着诱导时间的增加,红色荧光的酵母细胞所占比例不断增加,各组之间差异显著(P<0.05)。流式细胞仪检测结果显示,酵母细胞的荧光强度与诱导时间呈正比,其中诱导48h与72h的酵母细胞荧光强度不存在显著差异,基本趋于稳定不变的状态。因此,选取诱导48h为酵母表面展示的最佳诱导时间。上述研究结果表明SVCV的G蛋白已经成功展示于酿酒酵母细胞表面,研究为鲤春病毒血症酵母口服疫苗的研发奠定了前期基础。展开更多
The purpose of this study was to establish a method for the rapid detection of infectious pancreatic necrosis virus(IPNV,Jasper serotype)using reverse transcription loop-mediated isothermal amplification(RT-LAMP).Four...The purpose of this study was to establish a method for the rapid detection of infectious pancreatic necrosis virus(IPNV,Jasper serotype)using reverse transcription loop-mediated isothermal amplification(RT-LAMP).Four groups of specific primers were designed,according to the genome sequence of a Chinese IPNV isolate ChRtm213.The results showed that primer set B2 had the best amplification effect.When the final concentration of Mg2+was 6 mmol·L-1,dNTPs were 1 mmol·L-1 and betaine was 0.4 mol·L-1,the reaction could be completed in a 63℃water bath within 60 min.This RT-LAMP assay for the detection of IPNV had no cross-reactivity with infectious hematopoietic necrosis virus,viral hemorrhagic septicemia virus,grass carp reovirus and spring viremia of carp virus.The detection limit was 3.2×10-12 ng·μL-1.The sensitivity of this method was 10-fold higher than that of a previously published RT-LAMP assay for detecting the Spajarup(Sp)serotype of IPNV.This method,aimed at detecting IPNV isolates that were currently prevalent in China,possessed the characteristics of strong specificity,high sensitivity and direct interpretation by the naked eyes.The IPNV RT-LAMP was successfully applied to determine the clinical samples,which indicated the IPNV RT-LAMP assay was suitable for the rapid and large-scale detections of IPNV in China.展开更多
文摘鲤春病毒血症(spring virernia of carp,SVC)是由鲤春病毒血症病毒(carp spring virernia virus, CSVV)引起的多种鲤科鱼类高死亡率的病毒病,一般发生在春季,暴发后世界渔业经济损失严重。SVC是世界动物卫生组织(Office International Des Epizooties,OIE)必报的重要疫病,也是我国2008年最新颁布的动物疫病病种名录中的一类动物疫病。本文阐述了CSVV的病原生物学特性、流行病学、病理学,介绍国内外CSVV的检测技术手段、防治方法及疫苗的研发等,为CSVV的防治提供参考。
基金Supported by the National Natural Science Foundation of China(31802345)China Postdoctoral Science Foundation(2018M630893)Heilongjiang Province Postdoctoral Science Foundation(LBH-Z18275)。
文摘The purpose of this study was to establish a method for the rapid detection of infectious pancreatic necrosis virus(IPNV,Jasper serotype)using reverse transcription loop-mediated isothermal amplification(RT-LAMP).Four groups of specific primers were designed,according to the genome sequence of a Chinese IPNV isolate ChRtm213.The results showed that primer set B2 had the best amplification effect.When the final concentration of Mg2+was 6 mmol·L-1,dNTPs were 1 mmol·L-1 and betaine was 0.4 mol·L-1,the reaction could be completed in a 63℃water bath within 60 min.This RT-LAMP assay for the detection of IPNV had no cross-reactivity with infectious hematopoietic necrosis virus,viral hemorrhagic septicemia virus,grass carp reovirus and spring viremia of carp virus.The detection limit was 3.2×10-12 ng·μL-1.The sensitivity of this method was 10-fold higher than that of a previously published RT-LAMP assay for detecting the Spajarup(Sp)serotype of IPNV.This method,aimed at detecting IPNV isolates that were currently prevalent in China,possessed the characteristics of strong specificity,high sensitivity and direct interpretation by the naked eyes.The IPNV RT-LAMP was successfully applied to determine the clinical samples,which indicated the IPNV RT-LAMP assay was suitable for the rapid and large-scale detections of IPNV in China.
