Tilapia Lake Virus(TiLV)disease is an emerging and transboundary disease of tilapia cultures,causing mortality up to 90%globally.TiLV is a negative sense single stranded RNA virus belongs to family Amnoonviridae,genus...Tilapia Lake Virus(TiLV)disease is an emerging and transboundary disease of tilapia cultures,causing mortality up to 90%globally.TiLV is a negative sense single stranded RNA virus belongs to family Amnoonviridae,genus Tilapinevirus and species Tilapia tilapinevirus.The first TiLV outbreak to fishes was reported from Israel followed by other countries viz.,Ecuador,Colombia,Egypt,Thailand,Chinese Taipei,India,Malaysia,Bangladesh,Uganda,Tanzania,Peru,Mexico,Philippines,Indonesia,and USA.All the life stages of Tilapia(belonging to the family Cichlidae)are vulnerable to TiLV infection.However,river barb and giant gourami have also been found susceptible to TiLV infection.The virus infects the vital organs of the fish,including eyes,brain,and liver.The notable pathological finding of this disease includes syncytial cell formation and massive hepatocellular necrosis with pyknotic and karyolytic nuclei in the liver cells of infected fish.The disease is very contagious and spreads through both horizontal and vertical transmission.Several sensitive and rapid molecular diagnostic tools like reverse transcriptase polymerase chain reaction(RT-PCR),RT-quantitative PCR(RT-qPCR),loop-mediated isothermal amplification(LAMP)have been developed for early detection of the virus.Till date,no comprehensive control measures have been developed throughout the globe,although aggressive work on this line is going on.Implementations of strict good management practices,including quarantine protocols,are the only available option to combat the outbreak and spread of the disease.This review emphasizes the etiology,occurrence and distribution,mode of transmission,pathology and pathogenesis,diagnosis,possible control measures,and challenges of TiLV disease.展开更多
通过对NCBI数据库里的罗湖病毒(tilapia lake virus,TiLV)核苷酸序列进行生物信息学分析,了解罗湖病毒在全球的流行情况和中国罗非鱼(tilapia)养殖中罗湖病毒的感染情况。该研究将第3个片段的序列比对结果中的罗湖病毒分成12个簇群,通...通过对NCBI数据库里的罗湖病毒(tilapia lake virus,TiLV)核苷酸序列进行生物信息学分析,了解罗湖病毒在全球的流行情况和中国罗非鱼(tilapia)养殖中罗湖病毒的感染情况。该研究将第3个片段的序列比对结果中的罗湖病毒分成12个簇群,通过对片段进行motif序列富集,显示motif序列每个位点的碱基。对从广东各地区罗非鱼养殖场采集到的302份样本进行检测,结果显示,均未检测出罗湖病毒阳性,并且显示motif序列位点的碱基非常稳定。通过对罗湖病毒进行分子流行病学分析,中国虽未有罗湖病毒流行的趋势,但在进出口贸易中仍要严格防控罗湖病毒的输入,并应对这些非常保守的motif的相关功能进行深入研究。展开更多
【目的】制备抗罗非鱼湖病毒(Tilapia Lake Virus,TiLV)S10节段基因编码蛋白的单克隆抗体(Monoclonal antibody,MAb)。【方法】构建含有TiLV S10基因节段的重组表达质粒pET32a-S10,并将其转化至BL21(DE3)感受态细胞,经IPTG诱导表达和镍...【目的】制备抗罗非鱼湖病毒(Tilapia Lake Virus,TiLV)S10节段基因编码蛋白的单克隆抗体(Monoclonal antibody,MAb)。【方法】构建含有TiLV S10基因节段的重组表达质粒pET32a-S10,并将其转化至BL21(DE3)感受态细胞,经IPTG诱导表达和镍柱纯化获得高纯度重组S10蛋白;用纯化的重组蛋白免疫6~8周龄雌性BALB/c小鼠多次后,取其脾细胞与SP2/0细胞进行融合获得杂交瘤细胞,采用有限稀释法和ELISA方法,筛选获得2株能稳定分泌抗S10蛋白MAb的杂交瘤细胞株,分别命名为2C3和2E3,并对其特异性、亚型和效价进行分析。【结果】Western blot结果显示,2C3和2E3均能识别S10重组蛋白及TiLV,间接免疫荧光试验(IFA)结果表明,2C3和2E3只与TiLV感染的TiB细胞呈阳性反应。亚型检测结果显示,2C3抗体为IgG1/к型,2E3抗体为IgG2a/к型。抗体效价测定结果表明,两种MAb的效价分别为1∶12800,1∶51200。【结论】抗TiLV-S10蛋白MAb特异性强、效价高,可为后续TiLV疫苗的研发、免疫学方法的建立及S10蛋白功能的研究提供重要材料和支撑。展开更多
【目的】建立快速检测罗非鱼罗湖病毒(Tilapia Lake Virus,Ti LV)的RT-PCR方法。【方法】参照GenBank中罗非鱼罗湖病毒全基因序列,依据其假定蛋白基因片段3设计合成1对特异性扩增引物,提取罗湖病毒RNA,逆转录成cDNA,进行PCR扩增,通过优...【目的】建立快速检测罗非鱼罗湖病毒(Tilapia Lake Virus,Ti LV)的RT-PCR方法。【方法】参照GenBank中罗非鱼罗湖病毒全基因序列,依据其假定蛋白基因片段3设计合成1对特异性扩增引物,提取罗湖病毒RNA,逆转录成cDNA,进行PCR扩增,通过优化扩增条件和扩增体系,建立快速检测罗非鱼罗湖病毒的RT-PCR方法。用该方法对自然感染罗湖病毒发病的罗非鱼样品进行RT-PCR扩增。【结果】RT-PCR扩增得到与试验设计相符的499 bp的特异性目的条带,而对健康罗非鱼、野生罗非鱼及其他病毒对照组的扩增结果均为阴性。测序比对结果表明,该方法检测结果准确,最低可检测出约10fg/mL的质粒DNA,具有较好的特异性和较高的敏感性。用该方法对332份临床样品进行RT-PCR检测,其中36份样品扩增出特异性的目的条带,经测序比对,检测结果正确。【结论】建立的检测方法可用于TiLV的检测。展开更多
罗非鱼是世界上仅次于鲤科鱼的第二大养殖鱼类,是全球重要的动物蛋白食物来源。自2009年开始,由一种新型RNA病毒-罗非鱼湖病毒(Tilapia lake virus,TiLV)感染引起的罗非鱼湖病毒病(Tilapia lake virus disease,TiLVD),在全球各区域相继...罗非鱼是世界上仅次于鲤科鱼的第二大养殖鱼类,是全球重要的动物蛋白食物来源。自2009年开始,由一种新型RNA病毒-罗非鱼湖病毒(Tilapia lake virus,TiLV)感染引起的罗非鱼湖病毒病(Tilapia lake virus disease,TiLVD),在全球各区域相继暴发,造成了巨大经济损失,给全球的罗非鱼养殖业带来了巨大威胁和严峻挑战。TiLVD疫情已在全球多个不相连的区域均有发生,罗非鱼主养国家和区域都或多或少地受到TiLV的感染和威胁,TiLVD疫情有进一步扩大和加剧的趋势。目前,针对该疫病还没有任何有效的防控措施,但随着研究的深入和国际合作的加强,有望在早期精准诊断、传播途径阻断和疫苗免疫防控等方面实现突破。本文就TiLVD的病原学、流行病学、感染机制、诊断及防控等方面的研究进展进行综述,以期为TiLVD的基础研究和疫病防控提供参考。展开更多
基金the Dean,College of Fisheries,CAU(Imphal),Lembucherra,Tripura,799210,IndiaDirector ICAR-CIFA,Bhubaneswar,751002,India for providing the necessary facilities.
文摘Tilapia Lake Virus(TiLV)disease is an emerging and transboundary disease of tilapia cultures,causing mortality up to 90%globally.TiLV is a negative sense single stranded RNA virus belongs to family Amnoonviridae,genus Tilapinevirus and species Tilapia tilapinevirus.The first TiLV outbreak to fishes was reported from Israel followed by other countries viz.,Ecuador,Colombia,Egypt,Thailand,Chinese Taipei,India,Malaysia,Bangladesh,Uganda,Tanzania,Peru,Mexico,Philippines,Indonesia,and USA.All the life stages of Tilapia(belonging to the family Cichlidae)are vulnerable to TiLV infection.However,river barb and giant gourami have also been found susceptible to TiLV infection.The virus infects the vital organs of the fish,including eyes,brain,and liver.The notable pathological finding of this disease includes syncytial cell formation and massive hepatocellular necrosis with pyknotic and karyolytic nuclei in the liver cells of infected fish.The disease is very contagious and spreads through both horizontal and vertical transmission.Several sensitive and rapid molecular diagnostic tools like reverse transcriptase polymerase chain reaction(RT-PCR),RT-quantitative PCR(RT-qPCR),loop-mediated isothermal amplification(LAMP)have been developed for early detection of the virus.Till date,no comprehensive control measures have been developed throughout the globe,although aggressive work on this line is going on.Implementations of strict good management practices,including quarantine protocols,are the only available option to combat the outbreak and spread of the disease.This review emphasizes the etiology,occurrence and distribution,mode of transmission,pathology and pathogenesis,diagnosis,possible control measures,and challenges of TiLV disease.
文摘通过对NCBI数据库里的罗湖病毒(tilapia lake virus,TiLV)核苷酸序列进行生物信息学分析,了解罗湖病毒在全球的流行情况和中国罗非鱼(tilapia)养殖中罗湖病毒的感染情况。该研究将第3个片段的序列比对结果中的罗湖病毒分成12个簇群,通过对片段进行motif序列富集,显示motif序列每个位点的碱基。对从广东各地区罗非鱼养殖场采集到的302份样本进行检测,结果显示,均未检测出罗湖病毒阳性,并且显示motif序列位点的碱基非常稳定。通过对罗湖病毒进行分子流行病学分析,中国虽未有罗湖病毒流行的趋势,但在进出口贸易中仍要严格防控罗湖病毒的输入,并应对这些非常保守的motif的相关功能进行深入研究。
文摘【目的】建立快速检测罗非鱼罗湖病毒(Tilapia Lake Virus,Ti LV)的RT-PCR方法。【方法】参照GenBank中罗非鱼罗湖病毒全基因序列,依据其假定蛋白基因片段3设计合成1对特异性扩增引物,提取罗湖病毒RNA,逆转录成cDNA,进行PCR扩增,通过优化扩增条件和扩增体系,建立快速检测罗非鱼罗湖病毒的RT-PCR方法。用该方法对自然感染罗湖病毒发病的罗非鱼样品进行RT-PCR扩增。【结果】RT-PCR扩增得到与试验设计相符的499 bp的特异性目的条带,而对健康罗非鱼、野生罗非鱼及其他病毒对照组的扩增结果均为阴性。测序比对结果表明,该方法检测结果准确,最低可检测出约10fg/mL的质粒DNA,具有较好的特异性和较高的敏感性。用该方法对332份临床样品进行RT-PCR检测,其中36份样品扩增出特异性的目的条带,经测序比对,检测结果正确。【结论】建立的检测方法可用于TiLV的检测。
文摘罗非鱼是世界上仅次于鲤科鱼的第二大养殖鱼类,是全球重要的动物蛋白食物来源。自2009年开始,由一种新型RNA病毒-罗非鱼湖病毒(Tilapia lake virus,TiLV)感染引起的罗非鱼湖病毒病(Tilapia lake virus disease,TiLVD),在全球各区域相继暴发,造成了巨大经济损失,给全球的罗非鱼养殖业带来了巨大威胁和严峻挑战。TiLVD疫情已在全球多个不相连的区域均有发生,罗非鱼主养国家和区域都或多或少地受到TiLV的感染和威胁,TiLVD疫情有进一步扩大和加剧的趋势。目前,针对该疫病还没有任何有效的防控措施,但随着研究的深入和国际合作的加强,有望在早期精准诊断、传播途径阻断和疫苗免疫防控等方面实现突破。本文就TiLVD的病原学、流行病学、感染机制、诊断及防控等方面的研究进展进行综述,以期为TiLVD的基础研究和疫病防控提供参考。
