Virus Inactivation

病毒灭活
  • 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
    背景:灭活卵巢副痘病毒(iPPVO)对先天免疫细胞具有很强的免疫调节作用,使它成为一个有吸引力的治疗候选人。然而,关于iPPVO诱导的免疫反应中涉及的信号通路知之甚少。
    方法:在本研究中,我们系统地分析了不同类型的树突状细胞(DC)如何对iPPVO(Zylexis,菌株D1710)通过流式细胞术和ELISA在BALB/c和C57BL/c小鼠中,并通过蛋白质印迹和蛋白质谱分析研究了哪种信号通路与DC活化有关。
    结果:我们证明了骨髓来源的常规DCs(BM-cDCs)和骨髓来源的浆细胞样DCs(BM-pDCs)成熟并分泌IFN-α/β,以响应Zylexis刺激两种小鼠品系。同样,Zylexis促进pDC分泌IL-12/23p40和TNF。然而,在BALB/c小鼠中诱导IL-12/23p40和cDC分泌TNF,但在C57BL/6小鼠中不诱导。分析潜在的信号通路显示,iPPVO诱导的cDCs成熟是TLR9独立的,而pDC的成熟部分依赖于TLR9途径。此外,在两种小鼠品系中,cDC产生促炎细胞因子和pDC分泌IFN-α/β部分依赖于TLR9途径。因此,其他信号通路似乎参与DCs对iPPVO的反应,由蛋白质分析支持。
    结论:我们的数据为iPPVO传感器的多样性及其在不同菌株和物种之间的不同影响提供了有用的见解。
    BACKGROUND: Inactivated parapoxvirus ovis (iPPVO) exerts strong immunomodulatory effects on innate immune cells, making it an attractive therapeutic candidate. However, little is known about the signaling pathways that are involved in iPPVO-induced immune responses.
    METHODS: In this study, we systematically analyzed how different types of dendritic cells (DCs) react to iPPVO (Zylexis, strain D1701) in both BALB/c and C57BL/6 mice by flow cytometry and ELISAs, and investigated which signaling pathway is related to DC activation by Western blotting and protein profiling.
    RESULTS: We demonstrated that bone marrow-derived conventional DCs (BM-cDCs) and bone marrow-derived plasmacytoid DCs (BM-pDCs) matured and secreted type I interferons in response to Zylexis stimulation in both mouse strains. Similarly, Zylexis promoted the secretion of IL-12/23p40 and TNF by pDCs. However, IL-12/23p40 and TNF secretion by cDCs were induced in BALB/c mice but not in C57BL/6 mice. Analyzing the underlying signaling pathways revealed that iPPVO-induced maturation of cDCs was Toll-like receptor 9 (TLR9) independent, while the maturation of pDCs partially depended on the TLR9 pathway. Moreover, the production of proinflammatory cytokines by cDCs and the secretion of IFN-α/β by pDCs partially depended on the TLR9 pathway in both mouse strains. Therefore, other signaling pathways seem to participate in the response of DCs to iPPVO, supported by protein profiling.
    CONCLUSIONS: Our data provide useful insights into the diversity of iPPVO sensors and their varying effects across different strains and species.
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  • 文章类型: Journal Article
    摘要最近,高致病性禽流感A(H5N1)爆发,携带进化枝2.3.4.4b血凝素(HA)基因,自2021年冬季以来在北美鸟类种群中普遍存在,据报道在美国的奶牛中。截至2024年5月24日,该病毒已影响9个州的63个奶牛群,并导致2人感染。该病毒在奶牛中引起不寻常的症状,包括牛奶产量的意外下降,和浓浓的初乳般的牛奶。值得注意的是,美国食品和药物管理局报告说,大约20%的检测零售牛奶样品含有H5N1病毒,来自牛群感染地区的阳性结果百分比更高。关于巴氏杀菌如何有效地灭活牛奶中的H5N1病毒的数据很少。因此,在这项研究中,我们评估了H5进化枝2.3.4.4b病毒的热稳定性,以及一种人类H3N2病毒和其他流感亚型病毒,包括H1、H3、H7、H9和H10亚型病毒。我们还评估了巴氏杀菌在灭活这些病毒中的有效性。我们发现禽类H3病毒表现出最高的热稳定性,而属于进化枝2.3.4.4b的H5N1病毒表现出中等的热稳定性。重要的是,我们的数据提供了直接证据,表明乳品公司使用的标准巴氏灭菌方法可有效灭活原料奶中所有测试的流感病毒亚型.我们的发现表明,热巴氏杀菌奶制品不会对消费者构成安全风险。
    Recently, an outbreak of highly pathogenic avian influenza A (H5N1), which carries the clade 2.3.4.4b hemagglutinin (HA) gene and has been prevalent among North American bird populations since the winter of 2021, was reported in dairy cows in the United States. As of 24 May 2024, the virus has affected 63 dairy herds across nine states and has resulted in two human infections. The virus causes unusual symptoms in dairy cows, including an unexpected drop in milk production, and thick colostrum-like milk. Notably, The US Food and Drug Administration reported that around 20% of tested retail milk samples contained H5N1 viruses, with a higher percentage of positive results from regions with infected cattle herds. Data are scant regarding how effectively pasteurization inactivates the H5N1 virus in milk. Therefore, in this study, we evaluated the thermal stability of the H5 clade 2.3.4.4b viruses, along with one human H3N2 virus and other influenza subtype viruses, including H1, H3, H7, H9, and H10 subtype viruses. We also assessed the effectiveness of pasteurization in inactivating these viruses. We found that the avian H3 virus exhibits the highest thermal stability, whereas the H5N1 viruses that belong to clade 2.3.4.4b display moderate thermal stability. Importantly, our data provide direct evidence that the standard pasteurization methods used by dairy companies are effective in inactivating all tested subtypes of influenza viruses in raw milk. Our findings indicate that thermally pasteurized milk products do not pose a safety risk to consumers.
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  • 文章类型: Journal Article
    photo-Fenton工艺对于去除病原体是有效的,它的低成本版本可以应用于资源贫乏的环境中。在这里,使用低浓度的氧化铁(赤铁矿和磁铁矿)和过硫酸盐(过氧单硫酸盐-PMS,和过氧二硫酸盐-PDS),对MS2噬菌体表现出优异的灭活性能。在细菌的存在下,在H2O2和PDS系统中,MS2失活受到抑制,但在涉及PMS的系统中得到促进。所有提议的系统对混合细菌和病毒的灭活效力均大于唯一细菌的灭活效力,显示潜在的实际应用。还研究了掺入腐殖酸的氧化铁介导光Fenton样过程的失活性能;除了涉及PMS的系统外,H2O2-和PDS参与系统的失活功效被抑制,但PDS参与系统仍可接受(<2h)。反应性物种探索实验表明,·OH是H2O2和PDS系统中的主要自由基,而1O2在PMS参与系统中起关键作用。总之,低浓度的血铁矿和磁铁矿介导的过硫酸盐辅助的光Fenton样系统可以用作在阳光充足的地区进行病毒灭活的光Fenton过程的替代方法,为发展中国家的饮用水处理提供了更多的可能性。
    The photo-Fenton process is effective for pathogen removal, and its low-cost versions can be applied in resource-poor contexts. Herein, a photo-Fenton-like system was proposed using low concentrations of iron oxides (hematite and magnetite) and persulfates (peroxymonosulfate - PMS, and peroxydisulfate - PDS), which exhibited excellent inactivation performance towards MS2 bacteriophages. In the presence of bacteria, MS2 inactivation was inhibited in H2O2 and PDS systems but promoted in PMS-involved systems. The inactivation efficacy of all the proposed systems for mixed bacteria and viruses was greater than that of the sole bacteria, showing potential practical applications. The inactivation performance of humic acid-incorporated iron oxides mediating photo-Fenton-like processes was also studied; except for the PMS-involved system, the inactivation efficacy of the H2O2- and PDS-involved systems was inhibited, but the PDS-involved system was still acceptable (< 2 h). Reactive species exploration experiments indicated that ·OH was the main radical in the H2O2 and PDS systems, whereas 1O2 played a key role in the PMS-involved system. In summary, hematite- and magnetite-mediated persulfate-assisted photo-Fenton-like systems at low concentrations can be used as alternatives to the photo-Fenton process for virus inactivation in sunny areas, providing more possibilities for point-of-use drinking water treatment in developing countries.
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  • 文章类型: Journal Article
    病毒的灵敏鉴定和有效灭活对于病毒感染的早期诊断和治疗以防止病毒在环境中二次传播的风险至关重要。在这里,我们使用抗体/链霉亲和素双重标记的聚苯乙烯纳米珠和生物素标记的具有过氧化物酶模拟活性的G-四链体/血红素DNA酶开发了一种新颖的两步荧光免疫测定法,用于灵敏定量和有效灭活活着的寨卡病毒(ZIKV)。双标记纳米珠可以通过E蛋白靶向与ZIKV特异性结合,同时积累DNA酶,导致AmplexRed指示剂的催化氧化,并产生增强的聚集诱导的发射荧光信号,检测限低至66.3PFU/mL,准确度为100%。此外,辐射后由氧化的AmplexRed原位产生的强大的活性氧可以完全杀死病毒。这种灵敏而高效的检测-灭活集成系统将扩展病毒诊断工具并降低病毒传播在环境中的风险。
    Sensitive identification and effective inactivation of the virus are paramount for the early diagnosis and treatment of viral infections to prevent the risk of secondary transmission of viruses in the environment. Herein, we developed a novel two-step fluorescence immunoassay using antibody/streptavidin dual-labeled polystyrene nanobeads and biotin-labeled G-quadruplex/hemin DNAzymes with peroxidase-mimicking activity for sensitive quantitation and efficient inactivation of living Zika virus (ZIKV). The dual-labeled nanobeads can specifically bind ZIKV through E protein targeting and simultaneously accumulate DNAzymes, leading to the catalytic oxidation of Amplex Red indicators and generation of intensified aggregation-induced emission fluorescence signals, with a detection limit down to 66.3 PFU/mL and 100% accuracy. Furthermore, robust reactive oxygen species generated in situ by oxidized Amplex Red upon irradiation can completely kill the virus. This sensitive and efficient detection-inactivation integrated system will expand the viral diagnostic tools and reduce the risk of virus transmission in the environment.
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  • 文章类型: Journal Article
    水消毒无疑是确保人类用水安全的关键一步,臭氧被广泛用作控制水中病原微生物的高效消毒剂。尽管已广泛报道了复杂水基质中臭氧效率的降低,基质的各个成分对臭氧灭活病毒的具体作用程度尚不清楚,需要有效的方法来预测各种因素的综合影响。在这项研究中,从模拟的角度系统地研究和评估了复杂的水基质对病毒臭氧灭活的解耦影响。引入“等效臭氧消耗速率常数”(k\')的概念来量化不同物种的影响,并根据k值建立了动力学模型,用于模拟复杂基质中的臭氧失活过程。确定了不同物种影响臭氧失活有效性的机制:1)竞争效应(k\'=105~107M-1s-1),包括有机物和还原性离子(SO32-,NO2-,andI-),哪些是抑制病毒灭活的最具影响力的物种;2)屏蔽效应(k'=103~104M-1s-1),包括Ca2+,Mg2+,和高岭土;3)不显著影响(k\'=0~1M-1s-1),包括Cl-,SO42-,NO3-,NH4+,和Br-;4)促进作用(k\'=-103M-1s-1),包括CO32-和HCO3-。利用该模型成功实现了臭氧消毒效率的预测和复杂水生基质下物种贡献的评估。本研究中开发的系统理解和方法为预测复杂基质下的臭氧失活效率提供了可靠的框架,以及在实际废水处理过程中准确确定消毒剂剂量和控制干扰因素的潜在工具。
    Water disinfection is undoubtedly regarded as a critical step in ensuring the water safety for human consumption, and ozone is widely used as a highly effective disinfectant for the control of pathogenic microorganisms in water. Although the diminished ozone efficiencies in complex water matrices have been widely reported, the specific extent to which individual components of matrix act on the virus inactivation by ozone remains unclear, and effective methodologies to predict the comprehensive effects of various factors are needed. In this study, the decoupled impact of the intricate water matrix on the ozone inactivation of viruses was systematically investigated and assessed from a simulative perspective. The concept of \"equivalent ozone depletion rate constant\" (k\') was introduced to quantify the influence of different species, and a kinetic model was developed based on the k\' values for simulating the ozone inactivation processes in complex matrix. The mechanisms through which diverse species influenced the ozone inactivation effectiveness were identified: 1) competition effects (k\' = 105∼107 M-1s-1), including organic matters and reductive ions (SO32-, NO2-, and I-), which were the most influential species inhibiting the virus inactivation; 2) shielding effects (k\' = 103∼104 M-1s-1), including Ca2+, Mg2+, and kaolin; 3) insignificant effects (k\' = 0∼1 M-1s-1), including Cl-, SO42-, NO3-, NH4+, and Br-; 4) promotion effects (k\' = ∼-103 M-1s-1), including CO32- and HCO3-. Prediction of ozone disinfection efficiency and evaluation of species contribution under complex aquatic matrices were successfully realized utilizing the model. The systematic understanding and methodologies developed in this research provide a reliable framework for predicting ozone inactivation efficiency under complex matrix, and a potential tool for accurate disinfectant dosage determination and interfering factors control in actual wastewater treatment processes.
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  • 文章类型: Journal Article
    从2009年开始,H1N1已经成为困扰全球人口的呼吸道疾病之一。同时,由于COVID-19的影响,人们普遍认为,防止病毒传播需要个人防护措施和公共场所的消毒。
    这项研究进行了两个实验。在课堂实验中,对6个UVC剂量测试点进行了校准,以测试每个测试点的UVC剂量是否符合IAV灭活标准以及达到标准所需的时间.在模拟课堂实验中,放置了七个由IAV制成的正方形幻灯片。机器人运动10分钟后,将辐照过的无菌正方形载玻片制成悬浮液,并注入鸡胚。培养鸡胚并进行IAVs测试。
    课堂实验表明,5个测试点可以达到使IAV失活的标准(3mJ/cm2),所需时间为80分钟,40分钟,15分钟,5min和10min。80分钟内不符合标准的测试点的UVC剂量仅为0.5mJ/cm2。模拟课堂实验结果表明,99.99%的IAV被停用。此外,这项研究同时建立了桌面控制组和椅子手臂控制组,两者都产生了相同的结果,表示IAVs的失活对数≥4log。
    研究表明,在1.8mW/cm2的辐照密度下,物体表面的IAV可以有效且快速地失活。同时,该研究提供了在教室环境中使用GXU机器人使IAV失活的可行性的证据。
    UNASSIGNED: Starting from 2009, H1N1 has been one of the respiratory diseases that afflict the global population. Concurrently, due to the influence of COVID-19, it has become widely accepted that preventing the virus\'s spread necessitates personal protection measures and disinfection in public spaces.
    UNASSIGNED: This study conducted two experiments. In the classroom experiment, six UVC dose test points were calibrated to test whether the UVC dose at each testing point met the standards for inactivating IAVs and the time required to meet the standards. In the simulated classroom experiment, seven square slides made of IAVs were placed. After 10 min of robot movement, irradiated sterile square slides were made into suspension and injected into chicken embryos. Cultivate chicken embryos and conduct IAVs testing.
    UNASSIGNED: Classroom experiment has shown that 5 testing points can meet the standards for inactivating IAVs(3 mJ/cm2), with a required time of 80 min, 40 min, 15 min, 5 min and 10 min. The UVC dose for testing points that do not meet the standards in 80 min is only 0.5 mJ/cm2. The simulation classroom experiment outcomes revealed that 99.99 % of IAVs were deactivated. Furthermore, this study established both a desktop control group and a chair arm control group, both of which yielded identical results, indicating an inactivation logarithm of IAVs≥4log.
    UNASSIGNED: The study presented that IAVs on the surface of an object can be effectively and rapidly deactivated at an irradiation density of 1.8 mW/cm2. Meanwhile, the study provides evidence of the feasibility of using the GXU robot to inactivate IAVs in a classroom environment.
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  • 文章类型: Journal Article
    化学因素对DIV1感染性的影响尚未完全获得。为了研究DIV1对强盐水的稳定性,pH值,和其他化学条件,我们使用临床健康的南美白对虾个体进行了生物测定。DIV1接种物暴露于各种化学条件下,通过肌肉注射测定DIV1的感染性。结果表明,DIV1在强盐水中失去了传染性,特别是在3mol/L的NaCl溶液中持续1小时。此外,当经历低于3.1或高于9.6的pH水平时,发现DIV1在1小时内失活。此外,TritonX-100和1%甲醛都证明了使DIV1失活的能力。这些结果为DIV1对某些化学因素的耐受性提供了有价值的见解,作为制定针对DIV1的生物安全措施的参考。
    The effects of chemical factors on the infectivity of DIV1 have not been fully accessed yet. In order to investigate the stability of DIV1 to strong brine, pH, and other chemical conditions, we conducted a bioassay using clinically healthy Penaeus vannamei individuals. DIV1 inoculum was exposed to various chemical conditions, and the infectivity of DIV1 was determined through intramuscular injection. The results showed that DIV1 lost its infectivity when exposed to strong brine, specifically in a 3 mol/L NaCl solution for a duration of 1 h. Moreover, DIV1 was found to be inactivated within 1 h when subjected to pH levels below 3.1 or above 9.6. Additionally, both Triton X-100 and 1 % formaldehyde demonstrated the ability to inactivate DIV1. These results provide valuable insights into the tolerance of DIV1 towards certain chemical factors, serving as a reference for the establishment of biosecurity measures against DIV1.
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  • 文章类型: English Abstract
    针对治疗性抗体的市场需求,上游细胞培养规模和抗体表达滴度均有显著提高,而下游净化工艺的生产效率相对落后,下游处理能力已成为限制抗体生产吞吐量的瓶颈。以单克隆抗体mab-X为实验材料,优化了细胞培养液和低pH病毒灭活池的辛酸(CA)沉淀工艺条件,并研究了使用CA处理去除聚集体和灭活病毒的两种应用。根据实验室规模的研究,我们进行了500升的放大研究,其中将CA添加到低pH病毒灭活池中进行沉淀,并对沉淀前后的产品质量和产量进行了检测和比较。我们发现CA沉淀在蛋白A亲和色谱之前和之后均显着减少了HCP残留和聚集体。在骨料穗研究中,CA沉淀除去约15%的聚集体。病毒减少研究显示在蛋白A纯化抗体的CA沉淀期间模型逆转录病毒的完全清除。在放大研究中,深度过滤收获,亲和层析,低pH病毒灭活,CA沉淀和深度过滤,依次进行阳离子交换层析。CA沉淀进程中的搅拌时光和搅拌速度显著影响CA沉淀效力。CA沉淀后,低pH病毒灭活溶液中HCP残留量下降了895倍。降水后,产品纯度和HCP残留符合单克隆抗体的质量标准。CA沉淀可以减少常规纯化方法中的色谱步骤。总之,下游工艺中的CA沉淀可以简化常规纯化工艺,完全符合mab-X的净化质量标准,并提高生产效率,降低生产成本。本研究结果可能促进CA沉淀法在单克隆抗体纯化中的应用,为解决当前净化工艺的瓶颈提供参考。
    In response to the market demand for therapeutic antibodies, the upstream cell culture scale and expression titer of antibodies have been significantly improved, while the production efficiency of downstream purification process is relatively fall behind, and the downstream processing capacity has become a bottleneck limiting antibody production throughput. Using monoclonal antibody mab-X as experimental material, we optimized the caprylic acid (CA) precipitation process conditions of cell culture fluid and low pH virus inactivation pool, and studied two applications of using CA treatment to remove aggregates and to inactivate virus. Based on the lab scale study, we carried out a 500 L scale-up study, where CA was added to the low pH virus inactivation pool for precipitation, and the product quality and yield before and after precipitation were detected and compared. We found that CA precipitation significantly reduced HCP residuals and aggregates both before and after protein A affinity chromatography. In the aggregate spike study, CA precipitation removed about 15% of the aggregates. A virus reduction study showed complete clearance of a model retrovirus during CA precipitation of protein A purified antibody. In the scale-up study, the depth filtration harvesting, affinity chromatography, low pH virus inactivation, CA precipitation and depth filtration, and cation exchange chromatography successively carried out. The mixing time and stirring speed in the CA precipitation process significantly affected the CA precipitation effect. After CA precipitation, the HCP residue in the low pH virus inactivation solution decreased 895 times. After precipitation, the product purity and HCP residual meet the quality criteria of monoclonal antibodies. CA precipitation can reduce the chromatography step in the conventional purification process. In conclusion, CA precipitation in the downstream process can simplify the conventional purification process, fully meet the purification quality criterion of mab-X, and improve production efficiency and reduce production costs. The results of this study may promote the application of CA precipitation in the purification of monoclonal antibodies, and provide a reference for solving the bottleneck of the current purification process.
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  • 文章类型: Published Erratum
    [这更正了文章DOI:10.3389/fbioe.202.952498。].
    [This corrects the article DOI: 10.3389/fbioe.2022.952498.].
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