Virus Inactivation

病毒灭活
  • 文章类型: Journal Article
    在化学工程和水处理领域,对病毒的研究,包括代理人,有据可查。通常,代用品用于研究病毒及其行为,因为它们可以在更安全的条件下大量生产,并且更容易处理。事实上,代理人允许研究对人类无传染性但具有一些与致病病毒相似的特性的微生物:结构,composition,形态学,和大小。人类诺如病毒,被认为是所有年龄组的流行病和散发性胃肠炎病例的主要原因,可能会被杜兰病毒模仿.这项工作的目的是研究(i)杜兰病毒和诺如病毒的超滤,以验证杜兰病毒可以在水处理过程中用作诺如病毒的替代品,以及(ii)诺如病毒的保留和替代作为水质的功能,以更好地了解后一种致病病毒的使用。超滤测试显示病毒RNA中显著的对数减少值(LRV):全球LRV约为2.5(即,基于初始和渗透平均浓度),平均LRV在2和6之间(即,考虑到滞留物中病毒浓度的增加,保留率),诺如病毒和替代杜兰病毒。由于膜内腔中的病毒聚集,对于较高的初始浓度(从101至107个基因组拷贝/mL),获得较高的减少率(从2至6个对数基因组拷贝)。杜兰病毒似乎是通过膜过程保留诺如病毒的良好替代品。
    In the field of chemical engineering and water treatment, the study of viruses, included surrogates, is well documented. Often, surrogates are used to study viruses and their behavior because they can be produced in larger quantities in safer conditions and are easier to handle. In fact, surrogates allow studying microorganisms which are non-infectious to humans but share some properties similar to pathogenic viruses: structure, composition, morphology, and size. Human noroviruses, recognized as the leading cause of epidemics and sporadic cases of gastroenteritis across all age groups, may be mimicked by the Tulane virus. The objectives of this work were to study (i) the ultrafiltration of Tulane virus and norovirus to validate that Tulane virus can be used as a surrogate for norovirus in water treatment process and (ii) the retention of norovirus and the surrogate as a function of water quality to better understand the use of the latter pathogenic viruses. Ultrafiltration tests showed significant logarithmic reduction values (LRV) in viral RNA: around 2.5 for global LRV (i.e., based on the initial and permeate average concentrations) and between 2 and 6 for average LRV (i.e., retention rate considering the increase of viral concentration in the retentate), both for norovirus and the surrogate Tulane virus. Higher reduction rates (from 2 to 6 log genome copies) are obtained for higher initial concentrations (from 101 to 107 genome copies per mL) due to virus aggregation in membrane lumen. Tulane virus appears to be a good surrogate for norovirus retention by membrane processes.
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  • 文章类型: Case Reports
    西尼罗河病毒(WNV)是黄病毒科日本脑炎(JE)血清复合物的成员。我们报告了在最近的日本脑炎病毒(JEV)疫苗接种后,在2018年9月至2019年11月期间,四名全血献血者和一名血浆献血者进行了WNV核酸检测(NAT)反应性捐赠。
    病例1和4在接受JEV疫苗1天后有反应性WNVNAT捐赠。病例2在接受JEV疫苗3天后具有反应性WNV捐赠。病例3在从亚利桑那州返回后3天和接受JEV疫苗后1天具有反应性WNVNAT捐赠。病例5在接受JEV疫苗的同一天有反应性WNV捐赠。
    WNV筛选使用罗氏cobasWNV核酸测试(NAT)(罗氏分子系统)。国家微生物学实验室(NML)对WNV反应性捐赠进行了参考测试。还分析了JEV疫苗稀释物。
    对于案例1、3和5,补充NAT对于WNV和JEV是阴性的。案例2对于两个JEVNAT靶标之一具有弱扩增曲线。案例4为JEVNAT阳性,WNVNAT-负极。对病例2、4和5的捐赠标本进行的血清学测试不支持近期或远程WNV感染。通过cobas和补充NAT检测JEV疫苗稀释液。
    我们建议在JEV疫苗接种后实施临时献血延期,如果筛选利用具有检测JE血清复合物其他成员能力的WNV测定。
    West Nile Virus (WNV) is a member of the Japanese Encephalitis (JE) serocomplex within the Flaviviridae family. We report four whole blood donors and one plasma donor with WNV nucleic acid test (NAT)-reactive donations between September 2018 and November 2019, following recent Japanese Encephalitis virus (JEV) vaccination.
    Cases 1 and 4 had reactive WNV NAT donations 1 day after receiving the JEV vaccine. Case 2 had a reactive WNV donation 3 days after receiving the JEV vaccine. Case 3 had a reactive WNV NAT donation 3 days after returning from Arizona and 1 day after receiving the JEV vaccine. Case 5 had a reactive WNV donation the same day as receiving the JEV vaccine.
    WNV screening used the Roche cobas WNV nucleic acid test (NAT) (Roche Molecular Systems). Reference testing on WNV-reactive donations was carried out by the National Microbiology Laboratory (NML). JEV vaccine dilutions were also analyzed.
    Supplemental NAT was negative for WNV and JEV for Cases 1, 3, and 5. Case 2 had a weak amplification curve for one of two JEV NAT targets. Case 4 was JEV NAT-positive, WNV NAT-negative. Serologic testing on donation specimens for Cases 2, 4, and 5 did not support recent or remote WNV infection. JEV vaccine dilutions were detected by both cobas and supplemental NAT.
    We recommend implementing a temporary blood donor deferral following a JEV vaccination, if screening utilizes a WNV assay with the capability of detecting other members of the JE serocomplex.
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  • 文章类型: Journal Article
    与在生物安全3级和4级防护设施中操作电子显微镜(EM)相关的挑战减缓了高后果病毒的低温EM研究的进展。我们在委内瑞拉马脑炎病毒(VEEV)毒株TC-83的案例研究中解决了这一差距。病毒的化学灭活可能会物理上扭曲结构,因此为了验证天然结构的保留,我们选择了VEEV毒株TC-83来开发这种方法,因为该病毒具有4.8µ分辨率的低温EM结构。在我们的方法中,扩增的VEEVTC-83通过30%蔗糖垫直接从上清液中浓缩,重新暂停,并用1%戊二醛化学灭活。第二个30%蔗糖垫去除了任何可能干扰单颗粒分析的过量戊二醛。固定的低温电磁成像图,灭活的VEEV的分辨率为7.9µ。该图谱保留了天然病毒的结构特征,如二十面体对称,衣壳核心和三聚体尖峰的组织。我们的结果表明,我们的策略可以很容易地适应其他被包裹的失活,需要BSL-3或BSL-4用于冷冻EM的RNA病毒。然而,灭活的验证需要每个机构的生物安全委员会的监督。
    The challenges associated with operating electron microscopes (EM) in biosafety level 3 and 4 containment facilities have slowed progress of cryo-EM studies of high consequence viruses. We address this gap in a case study of Venezuelan Equine Encephalitis Virus (VEEV) strain TC-83. Chemical inactivation of viruses may physically distort structure, and hence to verify retention of native structure, we selected VEEV strain TC-83 to develop this methodology as this virus has a 4.8 Å resolution cryo-EM structure. In our method, amplified VEEV TC-83 was concentrated directly from supernatant through a 30 % sucrose cushion, resuspended, and chemically inactivated with 1 % glutaraldehyde. A second 30 % sucrose cushion removed any excess glutaraldehyde that might interfere with single particle analyses. A cryo-EM map of fixed, inactivated VEEV was determined to a resolution of 7.9 Å. The map retained structural features of the native virus such as the icosahedral symmetry, and the organization of the capsid core and the trimeric spikes. Our results suggest that our strategy can easily be adapted for inactivation of other enveloped, RNA viruses requiring BSL-3 or BSL-4 for cryo-EM. However, the validation of inactivation requires the oversight of Biosafety Committee for each Institution.
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  • 文章类型: Journal Article
    这项工作证明了如何将一系列工业批量分离步骤一起用于形成集成的自主下游过程的概念。在这个案例研究中的顺序包括阴离子色谱步骤,病毒灭活,最后是疏水色谱步骤。从批量分离到集成分离可最大限度地减少滞留时间,储罐,和所需的设备。从批量模式到集成模式的转换是通过从批量色谱中提取操作点和分离数据来实现的。集成的分离过程是在一个名为Orbit的开放式研究软件控制下实现的。使得可以操作包括多个步骤的复杂工艺配置。本案例研究的结果是自动化步骤的原理和方法,实现更连续和高效的下游过程。
    This work is a proof of concept of how a sequence of industrial batch separation steps together are used to form an integrated autonomous downstream process. The sequence in this case study consisted of an anion chromatography step, virus inactivation and finally a hydrophobic chromatography step. Moving from batch to integrated separation minimizes hold-up times, storage tanks, and required equipment. The conversion from batch to integrated mode is achieved by extracting operating points and separation data from batch chromatograms. The integrated separation process is realized on an ÄKTA Pure controlled by an open research software called Orbit, making it possible to operate complex process configurations including multiple steps. The results from this case study is the principle and method of the steps taken to automation, achieving a more continuous and efficient downstream process.
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  • 文章类型: Journal Article
    Arbovirus infections are increasing in prevalence worldwide. This presents new risks for blood transfusion. This article describes the epidemiology and surveillance of West Nile Virus, dengue and chikungunya and their role in the risk management of transfusions. Arboviruses are RNA viruses and very adaptable by nature. The majority of arbovirus infections are zoonoses. The risk of transmission is multifactorial and concerns the virus, vectors, animal reservoirs, the environment and human behaviour. In recent years, West Nile Virus has become established and widespread in North America, the number of cases of dengue worldwide has increased dramatically, and major epidemics of chikungunya have occurred in the Indian Ocean and Asia. The transmission of dengue and chikungunya is demonstrated in temperate zones. All arboviruses are potentially transmissible by transfusion due to their capacity to induce an asymptomatic viremic phase. The risk of West Nile Virus transmission via transfusion is recognised and prevention measures are well established. The risk of transmission via transfusion of dengue and chikungunya is real but difficult to quantify and the optimum prevention strategy is currently the subject of research. Access to up-to-date epidemiological data is an essential aid to decision-making, especially for donors returning from endemic areas to Europe. The challenge is to define and implement appropriate measures in unpredictable situations.
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  • 文章类型: Comment
    暂无摘要。
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  • 文章类型: Comparative Study
    A probabilistic model was used to compare cryoprecipitate to viral inactivated, commercial fibrinogen concentrate to evaluate with regard to the recipient\'s risk of exposure to an emergent AIDS-like epidemic. In patients who occasionally need a therapeutic dose of fibrinogen, commercial fibrinogen would be marginally safer than cryoprecipitate if the new pathogen were sensitive to inactivation. But there is a potential high risk of exposure if the emerging agent withstands inactivation. In most of the analyzed scenarios, cryoprecipitate is safer than commercial fibrinogen as long as the odds that the new agent is sensitive to inactivation are lower than 1.000 to 1.
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