Name: detection of viruses from bioaerosols using anion exchange resin
Uploaded: 2018-08-22T14:45:00.000+00:00
Duration: 6 min 10 s
Description: Watch this Scientific Journal Video about Detection of Viruses from Bioaerosols Using Anion Exchange Resin at JoVE.com

这项工作得到了来自 CDC/NIOSH 高平原山间农业健康与安全中心 (5U54OH008085) 和科罗拉多生物科学发现评估补助金计划 (14 改编-16) 的资助。

1. 气溶胶室的设置 (见图 2)
<ol><li>
预加载液体 impingers 与20毫升0.01 米磷酸盐缓冲盐水, pH 值 7.5 (PBS)。
	<ol>
<li>加入0.5 克阴离子交换树脂, 悬浮在其中一个液体 impingers 的 PBS 内, 另一种液体冲击器用作控制。</li>
	</ol>
</li>
	<li>在气溶胶腔内平行放置液体 impingers, 使用夹持器, 面对喷雾器的气溶胶入口。 注: 有关其他详细信息, 请参见图 2 。</li>
	<li>在液体 impingers 附近共找到一个直接读取的气溶胶监测器, 以测量气溶胶的质量浓度 (mg/米3)。</li>
	<li>在液体 impingers 附近共同定位其他直接读仪器 (室内空气质量监测和热风速计), 以监测气溶胶室内的环境变量, 如温度、相对湿度和风速。</li>
	<li>在气溶胶室的拐角处运行轴流风机, 以方便气溶胶的混合。 注: 轴流风机以固定速度运行, 不可调节。</li>
</ol>2. 模型 Bioaerosolization 实验 (见图 3)
<ol><li>对磷酸盐缓冲盐水 (PBS) 进行连续、10倍稀释的 MS2 噬菌体和流感疫苗的浓度。 注: 在这里演示的方法, 103.5 FFU/毫升的 a 型流感病毒疫苗菌株和 105 PFU/毫升的 MS2 噬菌体使用。为了确定菌的效价, 噬菌体被传播到大肠杆菌HS (pFamp) R, 并列举如前所述的20。该商业疫苗含有已知数量的四只活减毒菌株, 两种流感病毒和两种 B 型流感病毒, 以荧光聚焦单位或 FFU 表示。</li>
	<li>在6射流碰撞雾化器的玻璃容器内制备菌, 在100毫升的 PBS 中创建所需的病毒粒子浓度悬浮液, 并在自定义气溶胶腔内安装。</li>
	<li>启动仪器测量气溶胶腔内的变量, 如温度、相对湿度、二氧化碳浓度和风速 (使用轴流风机产生)。使用直接读取的气溶胶监测仪跟踪气溶胶室内的质量浓度, 并在试验之间建立一致性。</li>
	<li>密封气溶胶室, 用高效过滤空气净化10分钟。</li>
	<li>将经过过滤的干燥空气输送到6.9 帕的雾化器上。</li>
	<li>在每个气溶胶室试验中产生病毒 bioaerosols (通过雾化器) 的靶浓度。在此演示中, 将生成浓度为5毫克/米3的 bioaerosols。</li>
	<li>一旦达到目标质量浓度的气溶胶, 停止雾化。</li>
	<li>对每一个已校准为12.5 升/分流量的液体冲击器操作一个空气取样泵. 在每次取样事件前后使用主流量标准进行校准, 以确保采样量的一致性。使用位于雾化器附近的高效过滤线提供稀释空气, 并在燃烧室内保持恒定的压力。</li>
	<li>积极取样气溶胶室大气40分钟的时间, 以达到每样样品500升的取样量。 注: 在12.5 升/分两泵平行取样40分钟, 可取样气溶胶的1000升。因此, 在实验完成后, 预计气溶胶室中的大部分空气通过一个高效过滤系统被取样并释放到环境中。这是由颗粒浓度下降的证明。后实验, 表面被净化与10% 家用漂白剂和70% 乙醇。在本示范中使用指标;然而, 如果已知的病原体在该系统中使用, 则可根据需要实施额外的生物安全措施。</li>
</ol>3. 核酸提取和 qRT PCR 检测
<ol><li>
将核酸直接从阴离子交换树脂中分离出来, 并为比较起见, 从液体 impingers 中使用的液体隔离。在这个例子中, 我们描述了一个 RNA 隔离过程, 但是类似的协议可以用于 DNA 病毒。
	<ol>
<li>从负离子交换树脂中分离出核酸。<ol>
<li>将液体冲击器的阴离子交换树脂的所有液体转移到无菌50毫升圆锥管。</li>
			<li>允许树脂沉淀, 并慢慢地从负离子交换树脂中醒酒液体样品。使用1毫升吸管尖端, 以去除所有剩余的液体从阴离子交换树脂。</li>
			<li>从病毒 RNA 隔离试剂盒中, 将含有载体 RNA 的560µL 病毒裂解缓冲液直接添加到阴离子交换树脂中, 在室温下进行周期性混合孵化10分钟。</li>
			<li>使用1毫升吸管尖端, 转移病毒裂解物 (病毒裂解缓冲) 到一个无菌1.5 毫升圆锥管, 并进行 rna 隔离使用病毒 rna 隔离套件 (见材料表) 按照制造商的指示, 除了RNA 是洗脱在总容量60µL 洗脱的缓冲。</li>
		</ol>
</li>
		<li>从液体 impingers 中分离出核酸。<ol>
<li>吸管140µL 的液体样品成一个无菌1.5 毫升圆锥管和执行 rna 隔离使用病毒 rna 隔离套件按照制造商的指示, 除洗脱 RNA 的总容量60µL 洗脱缓冲。 注: 140 µL 是最大样本量, 可以处理与特定的病毒 RNA 隔离套件在这里使用的一个单一步骤的准备。</li>
		</ol>
</li>
	</ol>
</li>
	<li>执行 qRT PCR 检测 MS2 和流感 A 和 B 病毒, 如前所述23,24。在这一示范中, 5 µL 核酸 eluates 用于 qRT PCR。</li>
</ol>

<ol>
	<li>Pirtle, E. C., Beran, G. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Virus+survival+in+the+environment.">Virus survival in the environment.</a> Revue scientifique et technique (International Office of Epizootics). 10 (3), 733-748 (1991).</li><li>Nguyen, T. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+community-wide+outbreak+of+legionnaires+disease+linked+to+industrial+cooling+towers--how+far+can+contaminated+aerosols+spread?.">A community-wide outbreak of legionnaires disease linked to industrial cooling towers--how far can contaminated aerosols spread?.</a> The Journal of Infectious Diseases. 193 (1), 102-111 (2006).</li><li>Marks, P. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evidence+for+airborne+transmission+of+Norwalk-like+virus+(NLV)+in+a+hotel+restaurant.">Evidence for airborne transmission of Norwalk-like virus (NLV) in a hotel restaurant.</a> Epidemiology and Infection. 124 (3), 481-487 (2000).</li><li>Marks, P. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+school+outbreak+of+Norwalk-like+virus:+Evidence+for+airborne+transmission.">A school outbreak of Norwalk-like virus: Evidence for airborne transmission.</a> Epidemiology and Infection. 131 (1), 727-736 (2003).</li><li>Corzo, C. A., Culhane, M., Dee, S., Morrison, R. B., Torremorell, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Airborne+detection+and+quantification+of+swine+influenza+a+virus+in+air+samples+collected+inside,+outside+and+downwind+from+swine+barns.">Airborne detection and quantification of swine influenza a virus in air samples collected inside, outside and downwind from swine barns.</a> PLoS One. 8 (8), e71444 (2013).</li><li>Anderson, B. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bioaerosol+sampling+in+modern+agriculture:+A+novel+approach+for+emerging+pathogen+surveillance.">Bioaerosol sampling in modern agriculture: A novel approach for emerging pathogen surveillance.</a> The Journal of Infectious Diseases. 214 (4), 537-545 (2016).</li><li>Hietala, S. K., Hullinger, P. J., Crossley, B. M., Kinde, H., Ardans, A. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Environmental+air+sampling+to+detect+exotic+Newcastle+disease+virus+in+two+California+commercial+poultry+flocks.">Environmental air sampling to detect exotic Newcastle disease virus in two California commercial poultry flocks.</a> Journal of Veterinary Diagnostic Investigation. 17 (2), 198-200 (2005).</li><li>Jonges, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Wind-mediated+spread+of+low-pathogenic+avian+influenza+virus+into+the+environment+during+outbreaks+at+commercial+poultry+farms.">Wind-mediated spread of low-pathogenic avian influenza virus into the environment during outbreaks at commercial poultry farms.</a> PLoS One. 10 (5), e0125401 (2015).</li><li>Otake, S., Dee, S. A., Jacobson, L., Torremorell, M., Pijoan, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+aerosol+transmission+of+porcine+reproductive+and+respiratory+syndrome+virus+under+controlled+field+conditions.">Evaluation of aerosol transmission of porcine reproductive and respiratory syndrome virus under controlled field conditions.</a> The Veterinary Record. 150 (26), 804-808 (2002).</li><li>Wu, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aerosolized+avian+influenza+A+(H5N6)+virus+isolated+from+a+live+poultry+market,+China.">Aerosolized avian influenza A (H5N6) virus isolated from a live poultry market, China.</a> The Journal of Infection. 74 (1), 89-91 (2017).</li><li>Choi, M. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Live+animal+markets+in+Minnesota:+A+potential+source+for+emergence+of+novel+influenza+A+viruses+and+interspecies+transmission.">Live animal markets in Minnesota: A potential source for emergence of novel influenza A viruses and interspecies transmission.</a> Clinical Infectious Diseases. 61 (9), 1355-1362 (2015).</li><li>Haig, C. W., Mackay, W. G., Walker, J. T., Williams, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bioaerosol+sampling:+Sampling+mechanisms,+bioefficiency+and+field+studies.">Bioaerosol sampling: Sampling mechanisms, bioefficiency and field studies.</a> The Journal of Hospical Infection. 93 (3), 242-255 (2016).</li><li>Anderson, B. D., Lednicky, J. A., Torremorell, M., Gray, G. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+bioaerosol+aampling+for+airborne+virus+surveillance+in+swine+production+facilities:+A+mini+review.">The use of bioaerosol aampling for airborne virus surveillance in swine production facilities: A mini review.</a> Frontiers in Veterinary Science. 4, 121 (2017).</li><li>Verreault, D., Moineau, S., Duchaine, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methods+for+sampling+of+airborne+viruses.">Methods for sampling of airborne viruses.</a> Microbiology and Molecular Biology Reviews. 72 (3), 413-444 (2008).</li><li>Hogan, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sampling+methodologies+and+dosage+assessment+techniques+for+submicrometre+and+ultrafine+virus+aerosol+particles.">Sampling methodologies and dosage assessment techniques for submicrometre and ultrafine virus aerosol particles.</a> Journal of Applied Microbiology. 99 (6), 1422-1434 (2005).</li><li>Yu, K. -. P., Chen, Y. -. P., Gong, J. -. Y., Chen, Y. -. C., Cheng, C. -. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improving+the+collection+efficiency+of+the+liquid+impinger+for+ultrafine+particles+and+viral+aerosols+by+applying+granular+bed+filtration.">Improving the collection efficiency of the liquid impinger for ultrafine particles and viral aerosols by applying granular bed filtration.</a> Journal of Aerosol Science. 101, 133-143 (2016).</li><li>Perez-Mendez, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+a+simple+and+cost+effective+filter+paper-based+shipping+and+storage+medium+for+environmental+sampling+of+F-RNA+coliphages.">Evaluation of a simple and cost effective filter paper-based shipping and storage medium for environmental sampling of F-RNA coliphages.</a> J Virol Methods. 194 (1-2), 60-66 (2013).</li><li>Chandler, J. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Field-based+evaluation+of+a+male-specific+(F+)+RNA+coliphage+concentration+method.">Field-based evaluation of a male-specific (F+) RNA coliphage concentration method.</a> Journal of Virological Methods. 239, 9-16 (2017).</li><li>Perez-Mendez, A., Chandler, J. C., Bisha, B., Goodridge, L. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Concentration+of+enteric+viruses+from+tap+water+using+an+anion+exchange+resin-based+method.">Concentration of enteric viruses from tap water using an anion exchange resin-based method.</a> Journal of Virological Methods. 206, 95-98 (2014).</li><li>Perez-Mendez, A., Chandler, J. C., Bisha, B., Goodridge, L. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+an+anion+exchange+resin-based+method+for+concentration+of+F-RNA+coliphages+(enteric+virus+indicators)+from+water+samples.">Evaluation of an anion exchange resin-based method for concentration of F-RNA coliphages (enteric virus indicators) from water samples.</a> Journal of Virological Methods. 204, 109-115 (2014).</li><li>Kammerer, J., Carle, R., Kammerer, D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adsorption+and+ion+exchange:+Basic+principles+and+their+application+in+food+processing.">Adsorption and ion exchange: Basic principles and their application in food processing.</a> Journal of Agricultural and Food Chemistry. 59 (1), 22-42 (2011).</li><li>Chandler, J. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+method+for+the+improved+detection+of+aerosolized+influenza+viruses+and+the+male-specific+(F+)+RNA+coliphage+MS2.">A method for the improved detection of aerosolized influenza viruses and the male-specific (F+) RNA coliphage MS2.</a> Journal of Virological Methods. 246, 38-41 (2017).</li><li>Friedman, S. D., Cooper, E. M., Calci, K. R., Genthner, F. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Design+and+assessment+of+a+real+time+reverse+transcription-PCR+method+to+genotype+single-stranded+RNA+male-specific+coliphages+(Family+Leviviridae).">Design and assessment of a real time reverse transcription-PCR method to genotype single-stranded RNA male-specific coliphages (Family Leviviridae).</a> Journal of Virological Methods. 173 (2), 196-202 (2011).</li><li>Selvaraju, S. B., Selvarangan, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+three+influenza+A+and+B+real-time+reverse+transcription-PCR+assays+and+a+new+2009+H1N1+assay+for+detection+of+influenza+viruses.">Evaluation of three influenza A and B real-time reverse transcription-PCR assays and a new 2009 H1N1 assay for detection of influenza viruses.</a> Journal of Clinical Microbiology. 48 (11), 3870-3875 (2010).</li><li>Cademartiri, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immobilization+of+bacteriophages+on+modified+silica+particles.">Immobilization of bacteriophages on modified silica particles.</a> Biomaterials. 31 (7), 1904-1910 (2010).</li><li>Michen, B., Graule, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isoelectric+points+of+viruses.">Isoelectric points of viruses.</a> Journal of Appled Microbiology. 109 (2), 388-397 (2010).</li><li>Turgeon, N., Toulouse, M. J., Martel, B., Moineau, S., Duchaine, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+five+bacteriophages+as+models+for+viral+aerosol+studies.">Comparison of five bacteriophages as models for viral aerosol studies.</a> Applied and Environmental Microbiology. 80 (14), 4242-4250 (2014).</li><li>Vergara, G. G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+FRNA+coliphages+as+indicators+of+human+enteric+viruses+in+a+tropical+urban+freshwater+catchment.">Evaluation of FRNA coliphages as indicators of human enteric viruses in a tropical urban freshwater catchment.</a> Water Research. 79, 39-47 (2015).</li><li>Tung-Thompson, G., Libera, D. A., Koch, K. L., de Los Reyes, F. L., Jaykus, L. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aerosolization+of+a+human+norovirus+surrogate,+bacteriophage+MS2,+during+simulated+vomiting.">Aerosolization of a human norovirus surrogate, bacteriophage MS2, during simulated vomiting.</a> PLoS One. 10 (8), e0134277 (2015).</li></ol>

作者没有什么可透露的。

本协议概述了一种使用改进的液体 impingers 对 bioaerosols 进行敏感病毒捕获的方法。该方法对 bioaerosols 中病毒载量的检测和定量进行了优化。这里所展示的具体修改包括在普通液体冲击器中加入负离子交换树脂。该方法是为简化下游样品加工而开发的, 而其他样品处理技术, 如离心、过滤和沉淀法等, 并没有提供这样的优势。核酸的提取是直接从树脂, 不需要高容量 elutions 和多样品准备步骤, 最终有?...

该方法的目的是提供一个高度敏感的气溶胶取样平台, 以促进分子检测的负电荷病毒从 bioaerosols。微生物, 包括病毒微粒, 可以在 bioaerosols 中存活1长时间。Bioaerosols 可以旅行较远的距离和保持生存能力和传染性, 这证明了军团的疾病爆发的起源于距离6公里的工业冷却塔从受影响的人和导致18人死亡2。通过 bioaerosols 介导的病毒向人类间接传播可能发生在多种环境中, 并已证明诺如病毒在学校和餐馆3、4爆发。同样, 气溶胶传播病毒可能发生在农业环境中, 如在猪和家禽养殖场, 这一传输路线被认为是一个主要因素在病毒的移动之间的生产设施5,6,7,8,9。
对病毒携带的 bioaerosols 的有效取样可以改善快速诊断和预防疫情的准备, 如在中国活动物市场 bioaerosols 检测到 H5 流感病毒的示威活动中所示, 以及美国10,11。目前的气溶胶取样技术涉及许多不同的粒子捕获原理, 可大致分为 impingers、旋风、撞击和过滤器12。在本议定书范围之外, 详尽地涵盖了这些平台对 bioaerosols 病毒取样的所有利弊;但是, 可以说, 大多数这些取样装置尚未针对病毒和噬菌体13的收集进行优化。此外, 病毒微粒的传染性往往受到负面影响, 液体 impingers 认为更有效地保持病毒传染性比取样设备, 如固体撞击或过滤器14。然而, 液体撞击的一个缺点是靶稀释效应, 这是因为病毒在收集容器中相对较大的体积 (通常是≥20毫升) 收集的。另一个重要的缺点是液体 impingers 的次优效率, 浓缩颗粒 < 0.5 µM 大小15。然而, 这些装置的捕获效率可以通过固定在固体基质上来改善, 因为固定化可以增强病毒核酸的保存和病毒传染性16,17。
我们以前已经证明, 阴离子交换树脂是一种有效的工具, 从液体基质中捕获和浓缩病毒, 包括 F RNA 噬菌体, 甲型肝炎病毒, 人腺病毒, 和轮状病毒18,19 ,20。根据制造商的定义, 在这项工作中使用的阴离子交换树脂是一种吸附聚苯乙烯强碱性阴离子交换树脂, 功能性季胺基团介导在液体介质中对阴离子的吸引和捕获21.因此, 阴离子交换树脂有望捕获病毒的净负表面电荷, 包括许多肠道病毒, 流感病毒和其他与公共和动物健康有关的病毒。
目前的协议涉及增加负离子交换树脂的液体冲击器。在该系统中, 树脂作为冲击器液中捕获的病毒微粒的二次浓缩步骤。核酸可以直接洗脱在小体积, 提供了一个浓缩样本的分子分析。因此, 该方法的主要优点是提高了病毒检测灵敏度, 主要是通过减少样本量。另外, 由于对负电荷病毒的固有非特定捕获, 该方法可能适用于检测大量感兴趣的病毒。对 A 型和 B 型流感病毒的疫苗株和 FRNA 噬菌体 MS2 (MS2) 进行了验证。这些病毒随后使用标准的 qRT PCR 方法检测, 如前所述22。终端用户不应期望在执行此方法时遇到困难, 因为对当前现有设备的修改不构成对常规气溶胶取样和分析流程的重大干扰。

<table><tbody><tr><td>&lt;em&gt;Escherichia coli&lt;/em&gt; bacteriophage MS2 (ATCC 15597-B1)</td><td>American Type Culture Collection</td><td>ATCC 15597-B1</td><td></td></tr><tr><td>FluMist Quadrivalent</td><td>AstraZeneca</td><td>Contact manufacturer</td><td>Viral constitutents of this vaccine are subject to change on an annual basis</td></tr><tr><td>CFX96 Touch Real-Time PCR Detection System</td><td>Bio-Rad</td><td>1855195</td><td></td></tr><tr><td>Primers and probes</td><td>Integrated DNA Technologies</td><td>NA</td><td></td></tr><tr><td>0.2 &amp;micro;M sterile filter</td><td>NA</td><td>NA</td><td></td></tr><tr><td>1 L pyrex bottles or equivalent</td><td>NA</td><td>NA</td><td></td></tr><tr><td>1 mL pipet tips</td><td>NA</td><td>NA</td><td></td></tr><tr><td>1 mL pipettor</td><td>NA</td><td>NA</td><td></td></tr><tr><td>50 mL serological pipet</td><td>NA</td><td>NA</td><td></td></tr><tr><td>PCR tubes</td><td>NA</td><td>NA</td><td></td></tr><tr><td>Pipet-aid or equivalent</td><td>NA</td><td>NA</td><td></td></tr><tr><td>QIAamp Viral RNA Mini Kit</td><td>Qiagen</td><td>52904</td><td></td></tr><tr><td>QuantiTect Probe RT-PCR Kit</td><td>Qiagen</td><td>204443</td><td></td></tr><tr><td>Amberlite IRA-900 chloride form</td><td>Sigma-Aldrich</td><td>216585-500G</td><td></td></tr><tr><td>Phosphate buffered saline</td><td>Sigma-Aldrich</td><td>P5368-10PAK</td><td></td></tr><tr><td>Water (molecular biology grade)</td><td>Sigma-Aldrich</td><td>W4502-1L</td><td></td></tr><tr><td>Eppendorf DNA LoBind Microcentrifuge Tubes</td><td>ThermoFisher</td><td>13-698-791</td><td></td></tr><tr><td>Falcon 50 mL Conical Centrifuge Tubes&amp;nbsp;</td><td>ThermoFisher</td><td>14-432-22</td><td></td></tr><tr><td>Falcon Polypropylene Centrifuge Tubes</td><td>ThermoFisher</td><td>05-538-62</td><td></td></tr><tr><td>SuperScript III Platinum One-Step qRT-PCR Kit w/ROX</td><td>ThermoFisher</td><td>11745100</td><td></td></tr><tr><td>SKC Biosampler 20 mL, 3-piece glass set</td><td>SKC Inc.</td><td>225-9593</td><td></td></tr><tr><td>Vac-u-Go sample pumps</td><td>SKC Inc.</td><td>228-9695</td><td></td></tr><tr><td>Collison nebulizer (6-jet)</td><td>BGI Inc.</td><td>NA</td><td></td></tr><tr><td>HEPA capsule</td><td>PALL</td><td>12144</td><td></td></tr><tr><td>Q-TRAK indoor air quality monitor 8554</td><td>TSI Inc.</td><td>NA</td><td></td></tr><tr><td>Alnor velometer thermal anemometer AVM440-A</td><td>TSI Inc.</td><td>NA</td><td></td></tr><tr><td>SidePak AM510 personal aerosol monitor</td><td>TSI Inc.</td><td>NA</td><td></td></tr><tr><td>Bioaerosol chamber</td><td>NA</td><td>NA</td><td></td></tr></tbody></table>

detection of viruses from bioaerosols using anion exchange resin

该协议演示了一种针对病毒的自定义气溶胶采样方法。在该系统中, 负离子交换树脂与液体撞击式空气取样装置相结合, 有效地集中了 bioaerosols 的负电荷病毒。因此, 树脂在气溶胶取样工作流中充当额外的浓度步骤。然后直接从阴离子交换树脂中提取出病毒微粒的核酸, 得到的样品适用于分子分析。此外, 本议定书还描述了一个定制的气溶胶室, 能够在各种环境条件下产生带有病毒的 bioaerosols, 并允许对环境变量 (如温度、湿度) 进行连续监测,风速和气溶胶质量浓度。使用此协议的主要优点是增强了病毒检测的灵敏度, 这是通过与未修改的常规液体冲击器进行直接比较来评估的。其他优势包括可能集中各种负电荷病毒, 低成本的阴离子交换树脂 (每样0.14 美元), 并易于使用。缺点包括本议定书无法评估树脂吸附的病毒微粒的传染性, 并可能需要优化冲击器中使用的液体取样缓冲器。

图 1展示了基于电荷的从 bioaerosols 捕获病毒的原理, 通过将树脂加入液体基 impingers。图 2显示了自定义构建的气溶胶室的设置。图 3描述了设置 aerosolization 实验的步骤和确保质量控制的措施。图 4显示了从 bioaerosols 捕获的负电荷病毒 qRT PCR 检测的放大曲线。表 1显示了用?...

Watch this Scientific Journal Video about Detection of Viruses from Bioaerosols Using Anion Exchange Resin at JoVE.com

Detection of Viruses from Bioaerosols Using Anion Exchange Resin

证明了一种基于液体冲击的气溶胶病毒取样方法。当与下游分子检测相结合时, 该方法允许对 bioaerosols 的病毒进行简便、灵敏的检测。

用阴离子交换树脂检测 Bioaerosols 病毒

This protocol demonstrates a customized bioaerosol sampling method for viruses. In this system, anion exchange resin is coupled with liquid ...

detection-of-viruses-from-bioaerosols-using-anion-exchange-resin

Research

JoVE Journal

Environment

8.1K 次观看.  Colorado State University. 证明了一种基于液体冲击的气溶胶病毒取样方法。当与下游分子检测相结合时, 该方法允许对 bioaerosols 的病毒进行简便、灵敏的检测。

视频: Detection of Viruses from Bioaerosols Using Anion Exchange Resin

High-throughput Detection Method for Influenza Virus

Influenza virus is a respiratory pathogen that causes a high degree of morbidity and mortality every year in multiple parts of the world. Therefore, precise diagnosis of the infecting strain and rapid high-throughput screening of vast numbers of clinical samples is paramount to control the spread of pandemic infections. Current clinical diagnoses of influenza infections are based on serologic testing, polymerase chain reaction, direct specimen immunofluorescence and cell culture 1,2.
Here, we report the development of a novel diagnostic technique used to detect live influenza viruses. We used the mouse-adapted human A/PR/8/34 (PR8, H1N1) virus 3 to test the efficacy of this technique using MDCK cells 4. MDCK cells (104 or 5 x 103 per well) were cultured in 96- or 384-well plates, infected with PR8 and viral proteins were detected using anti-M2 followed by an IR dye-conjugated secondary antibody. M2 5 and hemagglutinin 1 are two major marker proteins used in many different diagnostic assays. Employing IR-dye-conjugated secondary antibodies minimized the autofluorescence associated with other fluorescent dyes. The use of anti-M2 antibody allowed us to use the antigen-specific fluorescence intensity as a direct metric of viral quantity. To enumerate the fluorescence intensity, we used the LI-COR Odyssey-based IR scanner. This system uses two channel laser-based IR detections to identify fluorophores and differentiate them from background noise. The first channel excites at 680 nm and emits at 700 nm to help quantify the background. The second channel detects fluorophores that excite at 780 nm and emit at 800 nm. Scanning of PR8-infected MDCK cells in the IR scanner indicated a viral titer-dependent bright fluorescence. A positive correlation of fluorescence intensity to virus titer starting from 102-105 PFU could be consistently observed. Minimal but detectable positivity consistently seen with 102-103 PFU PR8 viral titers demonstrated the high sensitivity of the near-IR dyes. The signal-to-noise ratio was determined by comparing the mock-infected or isotype antibody-treated MDCK cells.
Using the fluorescence intensities from 96- or 384-well plate formats, we constructed standard titration curves. In these calculations, the first variable is the viral titer while the second variable is the fluorescence intensity. Therefore, we used the exponential distribution to generate a curve-fit to determine the polynomial relationship between the viral titers and fluorescence intensities. Collectively, we conclude that IR dye-based protein detection system can help diagnose infecting viral strains and precisely enumerate the titer of the infecting pathogens.

This method describes the use of Infrared dye based imaging system for detection of H1N1 in bronchioalveolar lavage (BAL) fluid of infected mice at a high sensitivity. This methodology can be performed in a 96- or 384-well plate, requires &lt;10 &mu;l volume of test material and has the potential for concurrent screening of multiple pathogens.

流感病毒的高通量检测方法

Influenza virus is a respiratory pathogen that causes a high degree of morbidity and mortality every year in multiple parts of the world. Therefore, ...

科研

免疫与感染

Real-time Breath Analysis by Using Secondary Nanoelectrospray Ionization Coupled to High Resolution Mass Spectrometry

Exhaled volatile organic compounds (VOCs) have aroused considerable interest, since they can serve as biomarkers for disease diagnosis and environmental exposure in a non-invasive manner. In this work, we present a protocol to characterize the exhaled VOCs in real time by using secondary nanoelectrospray ionization coupled to high resolution mass spectrometry (Sec-nanoESI-HRMS). The homemade Sec-nanoESI source was readily set up based on a commercial nanoESI source. Hundreds of peaks were observed in the background-subtracted mass spectra of exhaled breath, and the mass accuracy values are -4.0-13.5 ppm and -20.3-1.3 ppm in the positive and negative ion detection modes, respectively. The peaks were assigned with accurate elemental composition according to the accurate mass and isotopic pattern. Less than 30 s is used for one exhalation measurement, and it takes approximately 7 min for six replicated measurements.

A protocol for characterizing chemical composition of exhaled breath in real time by using secondary nanoelectrospray ionization coupled to high resolution mass spectrometry is demonstrated.

利用二次 Nanoelectrospray 电离耦合高分辨质谱的实时呼吸分析

Exhaled volatile organic compounds (VOCs) have aroused considerable interest, since they can serve as biomarkers for disease diagnosis and environmental ...

化学

Capturing Actively Produced Microbial Volatile Organic Compounds from Human-Associated Samples with Vacuum-Assisted Sorbent Extraction

Volatile organic compounds (VOCs) from biological samples have unknown origins. VOCs may originate from the host or different organisms from within the host&#39;s microbial community. To disentangle the origin of microbial VOCs, volatile headspace analysis of bacterial mono- and co-cultures of Staphylococcus aureus, Pseudomonas aeruginosa, and Acinetobacter baumannii, and stable isotope probing in biological samples of feces, saliva, sewage, and sputum were performed. Mono- and co-cultures were used to identify volatile production from individual bacterial species or in combination with stable isotope probing to identify the active metabolism of microbes from the biological samples.
Vacuum-assisted sorbent extraction (VASE) was employed to extract the VOCs. VASE is an easy-to-use, commercialized, solvent-free headspace extraction method for semi-volatile and volatile compounds. The lack of solvents and the near-vacuum conditions used during extraction make developing a method relatively easy and fast when compared to other extraction options such as tert-butylation and solid phase microextraction. The workflow described here was used to identify specific volatile signatures from mono- and co-cultures. Furthermore, analysis of the stable isotope probing of human associated biological samples identified VOCs that were either commonly or uniquely produced. This paper presents the general workflow and experimental considerations of VASE in conjunction with stable isotope probing of live microbial cultures.

This protocol describes the extraction of volatile organic compounds from a biological sample with the vacuum-assisted sorbent extraction method, gas chromatography coupled with mass spectrometry using the Entech Sample Preparation Rail, and data analysis. It also describes culture of biological samples and stable isotope probing.

通过真空辅助吸附剂提取从人类相关样品中捕获活性产生的微生物挥发性有机化合物

Volatile organic compounds (VOCs) from biological samples have unknown origins. VOCs may originate from the host or different organisms from within the ...

生物化学

Rapid Molecular Detection and Differentiation of Influenza Viruses A and B

Influenza is a contagious respiratory illness caused by influenza viruses A and B in humans and causes a significant amount of morbidity and mortality every year. The Influenza A and B assay was the first CLIA-waived molecular rapid flu test available. The Influenza A and B test works by employing isothermal amplification with influenza-specific primers followed by target detection with molecular beacon probes. Here, the performance of the Influenza A and B assay on frozen, archived nasopharyngeal swab (NPS) specimens stored in viral transport medium (VTM) were compared to a respiratory panel assay.
The performance of the Influenza A and B assay was evaluated by comparing the results to the respiratory panel reference method. The sensitivity for total influenza virus A was 67.5% (95% CI (CI), 56.6-78.5) and the specificity was 86.9% (CI, 71.0-100). For influenza virus B testing, the sensitivity and specificity were 90.2% (CI, 68.5-100) and 98.8% (CI, 68.5-100), respectively.
This system has the advantage of a significantly shorter test time than any other currently available molecular assay and the simple, pipette-free procedure runs on a fully integrated, closed, small-footprint system. Overall, the Influenza A and B assay evaluated in this study has the potential to serve as a point-of-care rapid influenza diagnostic test.

We describe a rapid, molecular-based Influenza A and B assay. The Influenza assay detects each target within 15 min by employing isothermal amplification with influenza-specific primers followed by target detection with molecular beacon probes. The Influenza A and B assay is user-friendly and required minimal hands-on time to perform.

快速分子检测和流感病毒A和B的分化

Influenza is a contagious respiratory illness caused by influenza viruses A and B in humans and causes a significant amount of morbidity and mortality ...

High-throughput Detection of Respiratory Pathogens in Animal Specimens by Nanoscale PCR

Nanoliter scale real-time PCR uses spatial multiplexing to allow multiple assays to be run in parallel on a single plate without the typical drawbacks of combining reactions together. We designed and evaluated a panel based on this principle to rapidly identify the presence of common disease agents in dogs and horses with acute respiratory illness. This manuscript describes a nanoscale diagnostic PCR workflow for sample preparation, amplification, and analysis of target pathogen sequences, focusing on procedures that are different from microliter scale reactions. In the respiratory panel presented, 18 assays were each set up in triplicate, accommodating up to 48 samples per plate. A universal extraction and pre-amplification workflow was optimized for high-throughput sample preparation to accommodate multiple matrices and DNA and RNA based pathogens. Representative data are presented for one RNA target (influenza A matrix) and one DNA target (equine herpesvirus 1). The ability to quickly and accurately test for a comprehensive, syndrome-based group of pathogens is a valuable tool for improving efficiency and ergonomics of diagnostic testing and for acute respiratory disease diagnosis and management.

High-throughput testing of DNA and RNA based pathogens by nanoscale PCR is described using a syndromic canine and equine respiratory PCR panel.

在动物标本呼吸道病原体的高通量检测通过纳米PCR

Nanoliter scale real-time PCR uses spatial multiplexing to allow multiple assays to be run in parallel on a single plate without the typical drawbacks of ...

Engineering Antiviral Agents via Surface Plasmon Resonance

Surface plasmon resonance (SPR) is used to measure hemagglutinin (HA) binding to domain-swapped Cyanovirin-N (CV-N) dimer and to monitor interactions between mannosylated peptides and CV-N's high-affinity binding site. Virus envelope spikes gp120, HA, and Ebola glycoprotein (GP) 1,2 have been reported to bind both high- and low-affinity binding sites on dimeric CVN2. Dimannosylated HA peptide is also bound at the two low-affinity binding sites to an engineered molecule of CVN2, which is bearing a high-affinity site for the respective ligand and mutated to replace a stabilizing disulfide bond in the carbohydrate-binding pocket, thus confirming multivalent binding. HA binding is shown to one high-affinity binding site of pseudo-antibody CVN2 at a dissociation constant (KD) of 275 nM that further neutralizes human immunodeficiency virus type 1 (HIV-1) through oligomerization. Correlating the number of disulfide bridges in domain-swapped CVN2, which are decreased from 4 to 2 by substituting cystines into polar residue pairs of glutamic acid and arginine, results in reduced binding affinity to HA. Among the strongest interactions, Ebola GP1,2 is bound by CVN2 with two high-affinity binding sites in the lower nanomolar range using the envelope glycan without a transmembrane domain. In the present study, binding of the multispecific monomeric CV-N to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike (S) glycoprotein is measured at KD = 18.6 &#181;M as compared with nanomolar KD to those other virus spikes, and via its receptor-binding domain in the mid-&#181;-molar range.

Engineering Antiviral Agents via Surface Plasmon Resonance

The present protocol describes new tools for SPR binding assays to examine CV-N binding to HA, S glycoprotein, related hybrid-type glycans, and high-mannose oligosaccharides. SPR is used to determine the KD for binding either dimeric or monomeric CV-N to these glycans.

通过表面等离子体共振 设计 抗病毒药物

Surface plasmon resonance (SPR) is used to measure hemagglutinin (HA) binding to domain-swapped Cyanovirin-N (CV-N) dimer and to monitor interactions ...

生物工程

Remote Laboratory Management: Respiratory Virus Diagnostics

An uptick in recent pandemics (Ebola, Zika, MERS, influenza, etc.) underlines the need for a more 'nimble,' coordinated response that addresses a multitude of issues ranging from transportation, access, facilities, equipment, and communication to provider training. To address this need, we have developed an innovative, scalable, logistics-enhanced, mobile, laboratory facility for emergencies and epidemics in resource-constrained global settings. Utilizing a background in clinical operations as an academic medical center, we designed a rapidly-deployable, modular BSL-2 and BSL-3 facility with user-friendly software for tracking and management of drugs and supplies in remote regions during epidemics and outbreaks. Here, we present our intermodal, mobile, expandable shipping-container laboratory units. The design of the laboratory facilitates off-grid usage by minimizing power consumption and allowing alternate water sources. The unit's information communication technology (ICT) platform provides (i) user-friendly tablet-based documentation, (ii) enhanced tracking of patients and supplies, and (iii) integrated communication onsite with built-in telehealth capabilities. To ensure quality in remote environments, we have developed a checklist for a basic laboratory workflow and a protocol for respiratory viral diagnosis using reverse-transcription polymerase chain reaction (RT-PCR). As described, this innovative and comprehensive approach allows for the provision of laboratory capability in resource-limited global environments.

A rapidly-deployable, off-grid laboratory has been designed and built for remote, resource-constrained global settings. The features and critical aspects of the logistically-enhanced, expandable, multifunctional laboratory modules are explored. A checklist for a basic laboratory workflow and a protocol for a respiratory viral diagnostic test are developed and presented.

远程实验室管理: 呼吸道病毒诊断

An uptick in recent pandemics (Ebola, Zika, MERS, influenza, etc.) underlines the need for a more 'nimble,' coordinated response that addresses a ...

医学

Rapid and Specific Detection of Acinetobacter baumannii Infections Using a Recombinase Polymerase Amplification/Cas12a-based System

Acinetobacter baumannii, a gram-negative bacterium, is notorious for causing severe infections with high mortality rates. Rapid and accurate detection of A. baumannii is crucial for prompt treatment, effective infection control, and curbing antibiotic resistance. However, there is no suitable method for rapid and easy on-site detection of A. baumannii. The DNA Endonuclease Targeted CRISPR Trans Reporter (DETECTR) system offers a rapid, precise, and sensitive approach to A. baumannii detection by integrating the target-specific recognition capabilities of Cas12a with the isothermal amplification efficiency of Recombinase Polymerase Amplification (RPA). This protocol details the detection of A. baumannii using RPA combined with LbaCas12a endonuclease. The following steps are described in this article: extraction of DNA, selection of a specific DNA sequence, design of primer and CRISPR RNA (crRNA), construction of positive recombinant plasmid, setup of Cas12a-RPA assay, optimization of the RPA amplification system, visualization of the RPA-CRISPR/Cas12a assay using a fluorescence detection tool such as a real-time PCR instrument, and evaluation of sensitivity and specificity evaluation.

To facilitate the rapid and precise detection of Acinetobacter baumannii, we present a protocol that employs Recombinase Polymerase Amplification (RPA) in conjunction with LbaCas12a endonuclease for identifying A. baumannii infections.

Acinetobacter baumannii, a gram-negative bacterium, is notorious for causing severe infections with high mortality rates. Rapid and accurate detection of ...

Swabbing the Urban Environment - A Pipeline for Sampling and Detection of SARS-CoV-2 From Environmental Reservoirs

To control community transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) during the 2020 global pandemic, most countries implemented strategies based on direct human testing, face covering, and surface disinfection. Under the assumption that the main route of transmission includes aerosols and respiratory droplets, efforts to detect SARS-CoV-2 in fomites have focused on locations suspected of high prevalence (e.g., hospital wards, cruise ships, and mass transportation systems). To investigate the presence of SARS-CoV-2 on surfaces in the urban environment that are rarely cleaned and&#160;seldomly disinfected, 350 citizens were enlisted from the greater San Diego County. In total, these citizen scientists collected 4,080 samples. An online platform was developed to monitor sampling kit delivery and pickup, as well as to collect sample data. The sampling kits were mostly built from supplies available in pandemic-stressed stores. Samples were processed using reagents that were easy to access despite the recurrent supply shortage. The methods used were highly sensitive and resistant to inhibitors that are commonly present in environmental samples. The proposed experimental design and processing methods were successful at engaging numerous citizen scientists who effectively gathered samples from diverse surface areas. The workflow and methods described here are relevant to survey the urban environment for other viruses, which are of public health concern and pose a threat for&#160;future pandemics.

A citizen science project was designed to recruit San Diego residents to collect environmental samples for SARS-CoV-2. A multilingual web-based platform was created for data submission using a user-friendly mobile device interface. A laboratory information management system facilitated the collection of thousands of geographically diverse samples with real-time outcome tracking.

城市环境 -环境水库SARS-CoV-2采样和检测管道

To control community transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) during the 2020 global pandemic, most countries ...

生物学

用于呼吸道病毒检测的多重 RT-qPCR 检测

Development of Multiplex Real-Time RT-qPCR Assays for the Detection of SARS-CoV-2, Influenza A/B, and MERS-CoV

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that causes Coronavirus disease 2019 (COVID-19) is a serious threat to the general public&#39;s health. During influenza seasons, the spread of SARS-CoV-2 and other respiratory viruses may cause a population-wide burden of respiratory disease that is difficult to manage. For that, the respiratory viruses SARS-CoV-2, Influenza A, Influenza B, and Middle East respiratory syndrome (MERS-CoV) will need to be carefully watched over in the upcoming fall and winter seasons, particularly in the case of SARS-CoV-2, Influenza A, and Influenza B, which share similar epidemiological factors like susceptible populations, mode of transmission, and clinical syndromes. Without target-specific assays, it can be challenging to differentiate among cases of these viruses owing to their similarities. Accordingly, a sensitive and targeted multiplex assay that can easily differentiate between these viral targets will be useful for healthcare practitioners. In this study, we developed a real-time reverse transcriptase-PCR-based assay utilizing an in-house developed R3T one-step RT-qPCR kit for simultaneous detection of SARS-CoV-2, Influenza A, Influenza B, and SARS-CoV-2, MERS-CoV. With as few as 10 copies of their synthetic RNAs, we can successfully identify SARS-CoV-2, Influenza A, Influenza B, and MERS-CoV targets simultaneously with 100% specificity. This assay is found to be accurate, reliable, simple, sensitive, and specific. The developed method can be used as an optimized SARS-CoV-2, Influenza A, Influenza B, and SARS-CoV-2, MERS-CoV diagnostic assay in hospitals, medical centers, and diagnostic laboratories as well as for research purposes.

作者聚焦:呼吸道病毒多重检测的进展 

We present two probe-based in-house one-step RT-qPCR kits for common respiratory viruses. The first assay is for SARS-CoV-2 (N), Influenza A (H1N1 and H3N2), and Influenza B. The second is for SARS-Cov-2 (N) and MERS (UpE and ORF1a). These assays can be successfully implemented in any specialized laboratory.

开发用于检测SARS-CoV-2、甲型/乙型流感和MERS-CoV的多重实时RT-qPCR检测

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that causes Coronavirus disease 2019 (COVID-19) is a serious threat to the general ...

用阴离子交换树脂检测 Bioaerosols 病毒

摘要

探索更多视频

此视频中的章节

流感病毒的高通量检测方法

利用二次 Nanoelectrospray 电离耦合高分辨质谱的实时呼吸分析

通过真空辅助吸附剂提取从人类相关样品中捕获活性产生的微生物挥发性有机化合物

快速分子检测和流感病毒A和B的分化

在动物标本呼吸道病原体的高通量检测通过纳米PCR

通过表面等离子体共振设计抗病毒药物

远程实验室管理: 呼吸道病毒诊断

Rapid and Specific Detection of Acinetobacter baumannii Infections Using a Recombinase Polymerase Amplification/Cas12a-based System

城市环境 -环境水库SARS-CoV-2采样和检测管道

开发用于检测SARS-CoV-2、甲型/乙型流感和MERS-CoV的多重实时RT-qPCR检测

用阴离子交换树脂检测 Bioaerosols 病毒

摘要

探索更多视频

此视频中的章节

流感病毒的高通量检测方法

利用二次 Nanoelectrospray 电离耦合高分辨质谱的实时呼吸分析

通过真空辅助吸附剂提取从人类相关样品中捕获活性产生的微生物挥发性有机化合物

快速分子检测和流感病毒A和B的分化

在动物标本呼吸道病原体的高通量检测通过纳米PCR

通过表面等离子体共振 设计 抗病毒药物

远程实验室管理: 呼吸道病毒诊断

Rapid and Specific Detection of Acinetobacter baumannii Infections Using a Recombinase Polymerase Amplification/Cas12a-based System

城市环境 -环境水库SARS-CoV-2采样和检测管道

开发用于检测SARS-CoV-2、甲型/乙型流感和MERS-CoV的多重实时RT-qPCR检测

通过表面等离子体共振设计抗病毒药物