Name: detection of infectious virus from field-collected mosquitoes by vero cell culture assay
Uploaded: 2011-06-09T13:00:00
Description: Watch this Scientific Journal Video about Detection of Infectious Virus from Field-collected Mosquitoes by Vero Cell Culture Assay at JoVE.com

我们非常感谢博士的重要贡献。约翰安德森，安德鲁主要和Shirley蒂雷尔这项研究。这项工作是支持部分由从疾病控制和预防中心（U50/CCU116806-01-1）和美国农业部（58-6615-1-218，CONH00768，CONH00773）中心的拨款。

1。蚊子排序和识别<ol><li>在一个专门的生物安全2级（BSL - 2）的工作人员是受过训练的工作与生活的蚊子实验室，开放实验室长凳上执行下列程序。整个过程中保持一个“冷链”。 </li><li>麻醉住成蚊，灭蚊收集袋放置在-20℃冷冻5分钟。收集袋转移到一个绝缘容器干冰。关闭盖子，并揭露蚊子，以二氧化碳为1-3分钟，以确保完成击倒。 </li><li>到病床上的干冰置于预冷的泛塔麻醉蚊子。单独用细?...

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	<li>Chosewood, L. C., Wilson, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Biosafety in Microbiological and Biomedical Laboratories. , (2007).</li><li>Anderson, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+of+West+Nile+virus+from+mosquitoes,+crows,+and+a+Cooper's+hawk+in+Connecticut.">Isolation of West Nile virus from mosquitoes, crows, and a Cooper's hawk in Connecticut.</a> Science. 286, 2331-2333 (1999).</li><li>Armstrong, P. M., Andreadis, T. G., Anderson, J. F., Main, A. 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M., Andreadis, T. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Eastern+equine+encephalitis+virus+in+mosquitoes+and+their+role+as+bridge+vectors.">Eastern equine encephalitis virus in mosquitoes and their role as bridge vectors.</a> Emerg Infect Dis. 16, 1869-1874 (2010).</li><li>Andreadis, T. G., Anderson, J. F., Armstrong, P. M., Main, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolations+of+Jamestown+Canyon+virus+(Bunyaviridae:+Orthobunyavirus)+from+field-collected+mosquitoes+(Diptera:+Culicidae)+in+Connecticut,+USA:+a+ten-year+analysis,+1997-2006.">Isolations of Jamestown Canyon virus (Bunyaviridae: Orthobunyavirus) from field-collected mosquitoes (Diptera: Culicidae) in Connecticut, USA: a ten-year analysis, 1997-2006.</a> Vector Borne Zoonotic Dis. 8, 175-188 (2008).</li><li>Beaty, B. J., Calisher, C. H., Shope, R., Lennette, E. H., Lennette, D. A., Lennette, E. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chapter+11.">Chapter 11.</a> Diagnostic Procedures for Viral, Rickettsial, and Chlamydial Infections. , 189-212 (1995).</li><li>Karabatsos, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> International Catalogue of Arboviruses Including Certain Other Viruses of Vertebrates. , (1985).</li><li>Lanciotti, R. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+detection+of+west+nile+virus+from+human+clinical+specimens,+field-collected+mosquitoes,+and+avian+samples+by+a+TaqMan+reverse+transcriptase-PCR+assay.">Rapid detection of west nile virus from human clinical specimens, field-collected mosquitoes, and avian samples by a TaqMan reverse transcriptase-PCR assay.</a> J Clin Microbiol. 38, 4066-4071 (2000).</li><li>Lambert, A. J., Martin, D. A., Lanciotti, R. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Detection+of+North+American+eastern+and+western+equine+encephalitis+viruses+by+nucleic+acid+amplification+assays.">Detection of North American eastern and western equine encephalitis viruses by nucleic acid amplification assays.</a> J Clin Microbiol. 41, 379-385 (2003).</li></ol>

Vero细胞培养法作为一种有效的方法现场收集的病毒的多样性屏幕蚊子。这种专门针对一个或几个病毒的分子生物学方法的对比。此外，我们发现Vero细胞培养法，同样如果不是更多的7比RT - PCR检测的敏感，并为我们提供了为今后的研究中，储存在我们的参考收集的病毒株。然而，细胞培养系统可能无法在许多情况下，适当的。这种做法会带来额外的费用和培训，要求在BSL - 3实验室中生长?...

<table border="1"><tbody><tr><th>试剂/设备名称</th><th>公司</th><th>产品编号</th></tr><tr><td>胎牛血清</td><td> GIBCO - Invitrogen公司</td><td> 16140 </td></tr><tr><td> Anti-biotic/mycotic </td><td> GIBCO - Invitrogen公司</td><td> 15240 </td></tr><tr><td>碳酸氢钠碳酸氢钠，7.5％Soln。 </td><td> GIBCO - Invitrogen公司</td><td> 25080 </td></tr><tr><td> L -谷氨酰胺200MM 100X </td><td> GIBCO，Invitrogen公司</td><td> 25030 </td></tr><tr><td>粉最小必需培养基（MEM） </td><td> GIBCO - Invitrogen公司</td><td> 11700 </td></tr><tr><td> 10X贝科的PBS </td><td> GIBCO - Invitrogen公司</td><td> 14080 </td></tr><tr><td>胰蛋白酶EDTA </td><td> GIBCO - Invitrogen公司</td><td> 15400 </td></tr><tr><td>兔血清</td><td> GIBCO - Invitrogen公司</td><td> 16120 </td></tr><tr><td> Vero细胞（克隆E6） </td><td> ATCC </td><td> CRL - 1586 </td></tr><tr><td>小组织培养瓶中，通风帽，25厘米2 </td><td>猎鹰 - 碧迪</td><td> 353112 </td></tr><tr><td>大型组织培养瓶，通风帽，175厘米2 </td><td>猎鹰 - 碧迪</td><td> 353108 </td></tr><tr><td>镀铜BB的，4.5毫米</td><td> Crosman </td><td> 0767 </td></tr><tr><td>调音台轧机</td><td>瑞驰</td><td> MM300 </td></tr><tr><td> Isopack冷冻齿条</td><td> Eppendorf公司</td><td> 022510240 </td></tr></tbody></table>

detection of infectious virus from field-collected mosquitoes by vero cell culture assay

蚊子都传播了不同的病毒，包括重要的人类病原体，如西尼罗河病毒，登革热病毒，和chickungunya病毒的数量。许多这些病毒已加强在其流行范围扩大到新界，需要有效的监测和控制方案，以应对这些威胁。监控病毒活动的一个战略涉及从流行网站收集大量的蚊子和测试病毒感染。在这篇文章中，我们描述了如何处理的过程中，屏幕现场收集Vero细胞培养法对传染性病毒的蚊子。由陷阱的位置和物种的蚊子进行排序，并归纳为含≤50个人的池。汇集标本匀浆缓冲使用混频器厂和水相融合Vero细胞培养（克隆E6）到接种生理​​盐水。细胞培养监测3-7后接种在细胞培养中生长的任何病毒是通过适当的诊断方法确定天的细胞病变效应。利用这种方法，我们已经分离出9种不同的病毒，在美国康涅狄格州收集蚊子，其中，5个是已知的导致人类疾病。这三种病毒（波托西病毒，西尼罗河病毒，和拉克罗斯病毒）代表北美新英格兰地区自1999年以来的新纪录。能够检测到的病毒的广泛的多样性是至关重要的监测建立和新出现的病毒在蚊子种群。

Watch this Scientific Journal Video about Detection of Infectious Virus from Field-collected Mosquitoes by Vero Cell Culture Assay at JoVE.com

Detection of Infectious Virus from Field-collected Mosquitoes by Vero Cell Culture Assay

我们描述了一个方法来处理和屏幕现场收集到的蚊子的病毒Vero细胞培养法的多样性。通过采用这种技术的，我们已检测到4个分类家庭在康涅狄格州收集蚊子的9种不同的病毒。

从现场收集到的蚊子传染病毒Vero细胞培养法检测

Mosquitoes transmit a number of distinct viruses including important human pathogens such as West Nile virus, dengue virus, and chickungunya virus. Many ...

Research

JoVE Journal

Immunology and Infection

Live Cell Imaging of Alphaherpes Virus Anterograde Transport and Spread

顺行运输Alphaherpes病毒传播的活细胞成像

Advances in live cell fluorescence microscopy  techniques, as well as the construction of recombinant viral strains that  express fluorescent fusion ...

科研

免疫与感染

Organoids as Model for Infectious Diseases: Culture of Human and Murine Stomach Organoids and Microinjection of Helicobacter Pylori

类器官的模型传染病：人类的文化与小鼠胃组织体与幽门螺杆菌的显微注射

Recently infection biologists have employed stem cell derived cultures to answer the need for new and better models to study host-pathogen interactions. ...

Zika Virus Infectious Cell Culture System and the In Vitro Prophylactic Effect of Interferons

Zika Virus Infectious Cell Culture System and the In Vitro Prophylactic Effect of Interferons

兹卡病毒感染细胞培养系统和<em&gt;体外</em&gt;干扰素的预防作用

Zika Virus (ZIKV) is an emerging pathogen that is linked to fetal developmental abnormalities such as microcephaly, eye defects, and impaired growth. ZIKV ...

Rescue and Characterization of Recombinant Virus from a New World Zika Virus Infectious Clone

来自新世界Zika病毒感染性克隆的重组病毒的救援和表征

Infectious cDNA clones allow for genetic manipulation of a virus, thus facilitating work on vaccines, pathogenesis, replication, transmission and viral ...

Rapid, Safe, and Simple Manual Bedside Nucleic Acid Extraction for the Detection of Virus in Whole Blood Samples

快速、安全、简便的人工床边核酸提取法检测全血标本中的病毒

The rapid diagnosis of an infection is essential for the outbreak management, risk containment, and patient care. We have previously shown a method for ...

An Ex Vivo Chicken Primary Bursal-cell Culture Model to Study Infectious Bursal Disease Virus Pathogenesis

An Ex Vivo Chicken Primary Bursal-cell Culture Model to Study Infectious Bursal Disease Virus Pathogenesis

原发性鸡初囊-细胞培养模型研究传染性囊内病病毒发病机制

Infectious bursal disease virus (IBDV) is a birnavirus of economic importance to the poultry industry. The virus infects B cells, causing morbidity, ...

Identification of Mouse and Human Antibody Repertoires by Next-Generation Sequencing

近一代测序鉴定小鼠与人体抗体的分泌

The immense adaptability of antigen recognition by antibodies is the basis of the acquired immune system. Despite our understanding of the molecular ...

Human Serum Anti-aquaporin-4 Immunoglobulin G Detection by Cell-based Assay

人血清抗水药物蛋白-4 免疫球蛋白 g 的细胞检测

Anti-aquaporin-4 (AQP4) immunoglobulin G (IgG) is the core diagnostic biomarker for neuromyelitis optica spectrum disorders (NMOSD). The cell-based assay ...

Purification of High Yield Extracellular Vesicle Preparations Away from Virus

高产细胞外囊泡制剂的纯化远离病毒

One of the major hurdles in the field of extracellular vesicle (EV) research today is the ability to achieve purified EV preparations in a viral infection ...

Vector Competence Analyses on Aedes aegypti Mosquitoes using Zika Virus

Vector Competence Analyses on Aedes aegypti Mosquitoes using Zika Virus

使用寨卡病毒对 埃及伊 蚊的载体能力分析

The procedures presented describe a generalized methodology to infect Aedes aegypti mosquitoes with Zika virus under laboratory conditions to determine ...