Virus Inactivation

病毒灭活
  • 文章类型: Journal Article
    传染病是全球水产养殖业和保护计划损失的主要原因。同时,传染病对孵化场饲养和释放到自然栖息地进行保护的鱼类构成重大风险,包括五大湖湖st鱼(AcipenserFulvescens,即,GL-LST)。最近,同种疱疹病毒(湖鱼疱疹病毒2型,即,在两个成年GL-LST群体中检测到LSHV-2)能够在成年和少年GL-LST中诱导疾病和/或死亡。开始开发疾病预防和/或控制方法,体外实验旨在确定LSHV-2对孵化场和水产养殖设施中常用的消毒剂(Virkon®-Aquatic:过氧单硫酸钾;Ovadine®:聚乙烯吡咯烷酮碘复合物;和Perox-Aid®:过氧化氢)的敏感性。将培养的LSHV-2暴露于两种浓度的每种消毒剂(Virkon®-Aquatic:0.5%和1%;Ovadine®:50和100ppm;Perox-Aid®:500和1000ppm),一式两份,持续1、10和30分钟。曝光后,消毒剂被中和,在白st鱼×湖st鱼杂交细胞系(WSxLS)上经过14天的潜伏期后,通过比较有和没有消毒剂暴露的病毒的50%组织培养感染剂量(TCID50/mL)来计算减少百分比.当暴露于Perox-Aid®时,LSHV-2%的降低范围为58.7%至99.5%。当暴露于Ovadine®时,减少的百分比从99.4%到100%不等。最后,当暴露于Virkon®-Aquatic时,对于两种浓度和所有时间点,减少百分比为100%。本文的结果提供了Virkon®-Aquatic和Ovadine®对LSHV-2都是杀病毒的证据,并且可以代表在野外设置下降低病毒传播风险的手段。
    Infectious diseases are a leading cause of losses in the aquaculture industry and conservation programs globally. Simultaneously, infectious diseases pose a substantial risk to fish being hatchery-reared and released into natural habitats for conservation purposes, including the Great Lakes lake sturgeon (Acipenser fulvescens, i.e., GL-LST). Recently, an alloherpesvirus (lake sturgeon herpesvirus 2, i.e., LSHV-2) capable of inducing disease and/or mortality in adult and juvenile GL-LSTs was detected in two adult GL-LST populations. To begin developing disease prevention and/or control methods, in vitro experiments were designed to determine the susceptibility of LSHV-2 to disinfectants commonly used in hatchery and aquaculture facilities (Virkon®-Aquatic: potassium peroxymonosulfate; Ovadine®: polyvinylpyrrolidone iodine complex; and Perox-Aid®: hydrogen peroxide). Cultured LSHV-2 was exposed to each disinfectant at two concentrations (Virkon®-Aquatic: 0.5% and 1%; Ovadine®: 50 and 100 ppm; and Perox-Aid®: 500 and 1000 ppm) in duplicate for durations of 1, 10, and 30 min. Following exposure, the disinfectant was neutralized, and after a 14-day incubation period on a white sturgeon × lake sturgeon hybrid cell line (WSxLS), percent reduction was calculated by comparing the 50% tissue culture infectious doses (TCID50/mL) of the virus with and without disinfectant exposure. When exposed to Perox-Aid®, LSHV-2 percent reduction ranged from 58.7% to 99.5%. When exposed to Ovadine®, the percent reduction ranged from 99.4% to 100%. Lastly, the percent reduction when exposed to Virkon®-Aquatic was 100% for both concentrations and all timepoints. The results herein provide evidence that both Virkon®-Aquatic and Ovadine® are virucidal to LSHV-2 and may represent a means to reduce virus transmission risk under field settings.
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  • 文章类型: Journal Article
    病毒给全球造成了巨大的公共卫生负担,控制它们传播的关键因素之一是使用消毒剂灭活它们的能力。然而,由于固有的病毒特征(例如对常用灭活剂的顽抗)和外部因素(例如在应用灭活剂之前的不适当清洁,接触时间不当,等。).考虑到消毒剂应用不当的可能性(如接触时间短于推荐时间,消毒剂浓度不当,等。),了解消毒剂在有机负载存在下的性能很重要。要做到这一点,在研究消毒剂对不同病毒的功效时,经常使用模拟有机负荷的引入。然而,食源性病毒灭活研究中使用的不同类型的模拟有机载量及其对灭活的相对影响尚未进行审查。这篇综述的目的是调查用于研究食源性病毒灭活的不同模拟有机负荷制剂,以及介绍和比较这些不同制剂对病毒灭活的影响。这篇综述中的研究结果表明,许多模拟有机负荷制剂可以降低消毒剂对病毒的效力。根据这篇综述的发现,血,特别是血清或粪便,是许多测试中最常用和最有效的模拟有机负荷形式之一。
    Viruses impose a significant public health burden globally, and one of the key elements in controlling their transmission is the ability to inactivate them using disinfectants. However, numerous challenges to inactivating foodborne viruses exist due to inherent viral characteristics (such as recalcitrance to commonly used inactivation agents) and external factors (such as improper cleaning before application of inactivation agent, improper contact time, etc.). Given the potential for improper application of disinfectants (such as shorter than recommended contact time, improper disinfectant concentration, etc.), understanding the performance of a disinfectant in the presence of an organic load is important. To accomplish this, the introduction of simulated organic loads is often used when studying the efficacy of a disinfectant against different viruses. However, the different types of simulated organic loads used in foodborne virus inactivation studies or their relative effects on inactivation have not been reviewed. The purpose of this review is to survey different simulated organic load formulations used in studying foodborne virus inactivation, as well as present and compare the influence of these different formulations on viral inactivation. The findings included in this review suggest that many simulated organic load formulations can reduce disinfectants\' efficacy against viruses. Based on the findings in this review, blood, particularly serum or feces, are among the most commonly used and efficacious forms of simulated organic load in many tests.
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  • 文章类型: Journal Article
    我们调查了四种欧洲禽流感A(H5N1)病毒在全乳和半脱脂乳中的热稳定性,以及在美国当前的甲型流感(H5N1)奶牛爆发中它们在牛肾和肺细胞中的复制。结果显示热失活的应变依赖性差异,特别是在全脂牛奶中,和肺细胞中可变的复制功效。这些发现支持评估欧洲H5N1病毒在牛奶中的失活及其在牛细胞中的复制,协助生物安全协议和公共卫生措施。
    We investigated the thermostability of four European avian influenza A(H5N1) viruses in whole and semi-skimmed milk and their replication in bovine kidney and lung cells amid the current influenza A(H5N1) dairy cattle outbreak in the United States. Results showed strain-dependent differences in thermal inactivation, particularly in whole milk, and variable replication efficacy in lung cells. These findings support assessing the inactivation of European H5N1 viruses in milk and their replication in bovine cells, aiding biosafety protocols and public health measures.
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  • 文章类型: Journal Article
    裂谷热病毒(RVFV)是节肢动物传播的,人畜共患,出血热病毒,可在牲畜和人类中引起严重疾病。RVFV在以前被认为是非地方病的地区的传播,以及缺乏用于人类和动物的许可疫苗,在全球范围内构成了重大的健康和经济威胁。因此,在我们对这种病毒及其人畜共患病的理解和管理方面取得重大进展至关重要。RVFV被认为是生物恐怖主义病原体,and,因此,只有少数机构,设施,法律授权人员拘留和处理。此外,这种病毒必须在生物安全3级(BSL3)实验室按照严格的生物安全协议进行操作,以确保达到生物安全的最高标准。BSL2实验室只能处理某些减毒株,如MP12株,取决于所考虑的国家。为了帮助研究人员在最安全的条件下使用RVFV,本章介绍了有效的RVFV净化和灭活的有效方法。
    The Rift Valley fever virus (RVFV) is an arthropod-borne, zoonotic, hemorrhagic fever virus that can cause severe diseases both in livestock and humans. The spread of RVFV in areas previously considered as non-endemic together with the absence of licensed vaccines for use in humans and animals poses a major health and economic threat worldwide. It is therefore crucial to make major progresses in our understanding and management of this virus and its zoonosis. RVFV is considered a bioterrorism pathogen, and, thus, only a few institutes, facilities, and personnel are legally authorized to detain it and handle it. Moreover, this virus must be manipulated in a biosafety level 3 (BSL3) laboratory following strict biosafety protocols to ensure that biosecurity\'s highest standards are met. Only certain attenuated strains such as the MP12 strain can be handled in BSL2 laboratories, depending on the country considered. To assist researchers in working with RVFV in the safest possible conditions, this chapter presents validated methods for effective RVFV decontamination and inactivation.
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  • 文章类型: Journal Article
    由病原病毒引起的流行病对全球公共卫生构成严重威胁。电磁波是一种非接触和非电离辐射技术,已成为灭活细菌病原体的有效工具。在这项研究中,我们使用9.375GHz的电磁波研究了电磁波对致病性人类冠状病毒替代病毒MHV-A59的灭活效果和机制,并评估不同表面材料的失活效率。我们表明,9.375GHz电磁波通过破坏病毒颗粒使MHV-A59失活,信封或基因组。我们还发现,9.375GHz电磁波可以降低病毒在无生命材料如塑料表面的感染性,玻璃,布,和木头。总之,我们的结果表明,9.375GHz电磁波是一种有前途的消毒技术,可以防止病原病毒的传播和感染。
    Epidemics caused by pathogenic viruses are a severe threat to public health worldwide. Electromagnetic waves are a type of noncontact and nonionizing radiation technology that has emerged as an effective tool for inactivating bacterial pathogens. In this study, we used a 9.375 GHz electromagnetic wave to study the inactivation effect and mechanism of electromagnetic waves on MHV-A59, a substitute virus for pathogenic human coronavirus, and to evaluate the inactivation efficiency on different surface materials. We showed that 9.375 GHz electromagnetic waves inactivate MHV-A59 by destroying viral particles, envelopes, or genomes. We also found that 9.375 GHz electromagnetic waves can decrease the infectivity of viruses on the surface of inanimate materials such as plastic, glass, cloth, and wood. In conclusion, our results suggested that the 9.375 GHz electromagnetic wave is a promising disinfection technique for preventing the spread and infection of pathogenic viruses.
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  • 文章类型: Journal Article
    最近的COVID-19大流行引起了人们对有效空气消毒解决方案的兴趣。杀菌紫外线(GUV)辐射的应用是防止COVID-19空气传播以及其他现有和未来传染性空气传播疾病的绝佳竞争者。虽然GUV已被证明可有效灭活SARS-CoV-2,但有关紫外线敏感性和剂量要求的定量数据,需要预测和优化GUV解决方案的性能,仍然有限。在这项研究中,研究了雾化SARS-CoV-2对254nm紫外线(UV)辐射的紫外线敏感性。这是通过在配备有上层UV-C照明器的测试室中采用基于3D计算流体动力学的SARS-CoV-2灭活模拟并将结果与在同一测试室中执行的先前发布的测量进行比较来完成的。在这项研究中发现的紫外线敏感性为(0.6±0.2)m2/J,这相当于在3和6J/m2之间的D90剂量。这些值与基于文献中报道的其他电晕病毒和灭活数据的先前估计在相同的范围内。
    The recent COVID-19 pandemic has raised interest in efficient air disinfection solutions. The application of germicidal ultraviolet (GUV) irradiation is an excellent contender to prevent airborne transmission of COVID-19, as well as other existing and future infectious airborne diseases. While GUV has already been proven effective in inactivating SARS-CoV-2, quantitative data on UV susceptibility and dose requirements, needed to predict and optimize the performance of GUV solutions, is still limited. In this study, the UV susceptibility of aerosolized SARS-CoV-2 to 254 nm ultraviolet (UV) irradiation is investigated. This is done by employing 3D computational fluid dynamics based simulations of SARS-CoV-2 inactivation in a test chamber equipped with an upper-room UV-C luminaire and comparing the results to previously published measurements performed in the same test chamber. The UV susceptibility found in this study is (0.6 ± 0.2) m2/J, which is equivalent to a D90 dose between 3 and 6 J/m2. These values are in the same range as previous estimations based on other corona viruses and inactivation data reported in literature.
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  • 文章类型: Journal Article
    二氧化钛(TiO2)显示出作为自清洁材料的巨大潜力,可以灭活严重急性呼吸道综合症冠状病毒2(SARS-CoV-2)并防止病毒传播。这项研究提供了有关UV-A光对分子和原子水平上TiO2表面上吸附的SARS-CoV-2病毒样颗粒(VLP)的光催化失活的影响的见解。X射线光电子能谱,结合密度泛函理论计算,表明刺突蛋白可以主要通过其氨基酸嵌段中的胺和酰胺官能团吸附在TiO2上。我们使用原子力显微镜和掠入射小角X射线散射(GISAXS)研究了在光照下VLP在TiO2上失活过程中的分子尺度形态变化。值得注意的是,原位测量揭示了VLP的光诱导形态变化,导致颗粒直径增加。这些结果表明,由紫外线照射引起的结构蛋白的变性和通过光催化反应的病毒结构的氧化可以在TiO2表面上发生。在N2气氛下的原位GISAXS测量显示病毒形态在UV光下保持完整。这提供了氧和UV光的存在对于在表面上引发光催化反应并随后使吸附的病毒失活是必要的证据。在这项研究中获得的对病毒灭活过程的化学见解极大地有助于开发用于灭活包膜病毒的固体材料。
    Titanium dioxide (TiO2) shows significant potential as a self-cleaning material to inactivate severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and prevent virus transmission. This study provides insights into the impact of UV-A light on the photocatalytic inactivation of adsorbed SARS-CoV-2 virus-like particles (VLPs) on a TiO2 surface at the molecular and atomic levels. X-ray photoelectron spectroscopy, combined with density functional theory calculations, reveals that spike proteins can adsorb on TiO2 predominantly via their amine and amide functional groups in their amino acids blocks. We employ atomic force microscopy and grazing-incidence small-angle X-ray scattering (GISAXS) to investigate the molecular-scale morphological changes during the inactivation of VLPs on TiO2 under light irradiation. Notably, in situ measurements reveal photoinduced morphological changes of VLPs, resulting in increased particle diameters. These results suggest that the denaturation of structural proteins induced by UV irradiation and oxidation of the virus structure through photocatalytic reactions can take place on the TiO2 surface. The in situ GISAXS measurements under an N2 atmosphere reveal that the virus morphology remains intact under UV light. This provides evidence that the presence of both oxygen and UV light is necessary to initiate photocatalytic reactions on the surface and subsequently inactivate the adsorbed viruses. The chemical insights into the virus inactivation process obtained in this study contribute significantly to the development of solid materials for the inactivation of enveloped viruses.
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  • 文章类型: Journal Article
    人类诺如病毒(HuNoV)是一种肠道感染性病原体,属于卡利病毒科,偶尔会引起流行病。容易通过食源性途径传播的循环酒精耐受性病毒颗粒显著导致HuNoV诱导的胃肠炎的全球负担。此外,与环境中其他微生物分泌的酶接触会影响病毒的感染性。因此,了解杯状病毒科的循环动力学对于减轻流行病至关重要。因此,在这项研究中,我们筛选了环境是否丰富的分泌酶成分,特别是蛋白酶,影响杯状病毒科的感染性。结果表明,将芽孢杆菌的丝氨酸蛋白酶与链霉菌产生的ε-聚-L-赖氨酸(EPL)结合起来,包括针对疫情的HuNoVGII.4_Sydney_2012菌株。体外和体内生化和病毒学分析表明,EPL具有两种独特的协同病毒灭活功能。首先,其维持最佳pH以促进蛋白酶敏感结构的病毒表面构象变化。随后,其通过在VP1衣壳中的P2和S结构域处的部分蛋白酶消化来抑制病毒RNA基因组释放。这项研究提供了有关细菌和Caliciviridae之间的高维环境相互作用的新见解,同时促进以蛋白酶为基础的抗病毒消毒剂的发展。
    Human norovirus (HuNoV) is an enteric infectious pathogen belonging to the Caliciviridae family that causes occasional epidemics. Circulating alcohol-tolerant viral particles that are readily transmitted via food-borne routes significantly contribute to the global burden of HuNoV-induced gastroenteritis. Moreover, contact with enzymes secreted by other microorganisms in the environment can impact the infectivity of viruses. Hence, understanding the circulation dynamics of Caliciviridae is critical to mitigating epidemics. Accordingly, in this study, we screened whether environmentally abundant secretase components, particularly proteases, affect Caliciviridae infectivity. Results showed that combining Bacillaceae serine proteases with epsilon-poly-L-lysine (EPL) produced by Streptomyces-a natural antimicrobial-elicited anti-Caliciviridae properties, including against the epidemic HuNoV GII.4_Sydney_2012 strain. In vitro and in vivo biochemical and virological analyses revealed that EPL has two unique synergistic viral inactivation functions. First, it maintains an optimal pH to promote viral surface conformational changes to the protease-sensitive structure. Subsequently, it inhibits viral RNA genome release via partial protease digestion at the P2 and S domains in the VP1 capsid. This study provides new insights regarding the high-dimensional environmental interactions between bacteria and Caliciviridae, while promoting the development of protease-based anti-viral disinfectants.
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  • 文章类型: Journal Article
    单克隆抗体(mAb)是治疗癌症的重要治疗蛋白,自身免疫性和罕见疾病。在生产过程中,storage,和管理过程,这些蛋白质会遇到各种压力因素,如温度波动,振动,和曝光,能够诱导对其结构的化学物理修饰。病毒灭活是下游过程中的关键步骤,它是通过在低pH下滴定mAb来实现的,其次是中和。pH值的变化会导致蛋白质的解折叠和随后的聚集,从而影响他们的生产。这项研究旨在调查在病毒灭活过程中联合暴露于光线是否会进一步影响伊匹单抗的结构完整性。主要用于治疗转移性黑色素瘤的mAb。Ipilimumab的生物物理和生化表征表明,pH变化对其在pH2下不可逆解折叠的稳定性具有相当大的风险。Ipilimumab变性的阈值介于pH2和3之间,并且与蛋白质结构协同性的丧失相关,这是决定蛋白质重折叠的最关键因素。光已经证明会加剧一些局部和全球影响,使得pH值引起的暴露区域更容易受到结构和化学变化的影响。因此,在单克隆抗体的灭菌过程中,应考虑对现实生活中暴露于环境光的具体预防措施,以避免失去治疗活性并增加产量。我们的发现强调了pH优化在保持mAb的结构完整性和治疗功效中的关键作用。此外,对Ipilimumab结构修饰的详细构象研究可能会改善这种有效药物的化学物理知识,并为在某种应激条件下更稳定的产品提出新的生产策略.
    Monoclonal antibodies (mAbs) are an essential class of therapeutic proteins for the treatment of cancer, autoimmune and rare diseases. During their production, storage, and administration processes, these proteins encounter various stressors such as temperature fluctuations, vibrations, and light exposure, able to induce chemico-physical modifications to their structure. Viral inactivation is a key step in downstream processes, and it is achieved by titration of the mAb at low pH, followed by neutralization. The changes of the pH pose a significant risk of unfolding and subsequent aggregation to proteins, thereby affecting their manufacturing. This study aims to investigate whether a combined exposure to light during the viral inactivation process can further affect the structural integrity of Ipilimumab, a mAb primarily used in the treatment of metastatic melanoma. The biophysical and biochemical characterization of Ipilimumab revealed that pH variation is a considerable risk for its stability with irreversible unfolding at pH 2. The threshold for Ipilimumab denaturation lies between pH 2 and 3 and is correlated with the loss of the protein structural cooperativity, which is the most critical factor determining the protein refolding. Light has demonstrated to exacerbate some local and global effects making pH-induced exposed regions more vulnerable to structural and chemical changes. Therefore, specific precautions to real-life exposure to ambient light during the sterilization process of mAbs should be considered to avoid loss of the therapeutic activity and to increase the yield of production. Our findings underscore the critical role of pH optimization in preserving the structural integrity and therapeutic efficacy of mAbs. Moreover, a detailed conformational study on the structural modifications of Ipilimumab may improve the chemico-physical knowledge of this effective drug and suggest new production strategies for more stable products under some kind of stress conditions.
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  • 文章类型: Journal Article
    甲醛灭活的脊髓灰质炎病毒颗粒的电化学分析表明,D抗原浓度与脊髓灰质炎病毒样品的最大振幅电流强度之间存在关系。因此,所得信号被鉴定为脊髓灰质炎病毒表面蛋白的电化学氧化。使用衣壳蛋白氨基酸残基的电氧化注册,通过5kGy剂量加速的电子灭活的脊髓灰质炎病毒颗粒的比较电化学分析,10kGy,15kGy,25kGy,在室温下进行30kGy。辐射剂量的增加伴随着电氧化信号的增加。在15-30kGy剂量的照射下,检测到脊髓灰质炎病毒衣壳蛋白的电氧化信号显着增加。获得的数据表明,在脊髓灰质炎病毒灭活条件下,脊髓灰质炎病毒衣壳蛋白的谱变化和电氧化信号增加与表面蛋白结构重组程度的增加和D-抗原保存不足有关。
    Electrochemical profiling of formaldehyde-inactivated poliovirus particles demonstrated a relationship between the D-antigen concentration and the intensity of the maximum amplitude currents of the poliovirus samples. The resultant signal was therefore identified as electrochemical oxidation of the surface proteins of the poliovirus. Using registration of electrooxidation of amino acid residues of the capsid proteins, a comparative electrochemical analysis of poliovirus particles inactivated by electrons accelerated with doses of 5 kGy, 10 kGy, 15 kGy, 25 kGy, 30 kGy at room temperature was carried out. An increase in the radiation dose was accompanied by an increase in electrooxidation signals. A significant increase in the signals of electrooxidation of poliovirus capsid proteins was detected upon irradiation at doses of 15-30 kGy. The data obtained suggest that the change in the profile and increase in the electrooxidation signals of poliovirus capsid proteins are associated with an increase in the degree of structural reorganization of surface proteins and insufficient preservation of the D-antigen under these conditions of poliovirus inactivation.
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