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Rapid isothermal point-of-care test for screening of SARS-CoV-2 (COVID-19) 快速等温定点检测筛查SARS-CoV-2 (COVID-19)
Pub Date : 2023-01-01 DOI: 10.1016/j.amolm.2023.100002
Jean-Marc Zingg , Yu-Ping Yang , Spencer Seely , Pratibha Joshi , Md Harun Or Roshid , Fabiola Iribarren Latasa , Gregory O'Connor , Jennifer Alfaro , Eduardo Riquelme , Sebastian Bernales , Emre Dikici , Sapna Deo , Sylvia Daunert

Rapid on-site diagnosis of emerging pathogens is key for early identification of infected individuals and for prevention of further spreading in a population. Currently available molecular diagnostic tests are instrument-based whereas rapid antibody and antigen tests are often not sufficiently sensitive for detection in pre-symptomatic subjects. There is a need for rapid point of care molecular screening tests that can be easily adapted to emerging pathogens and are selective, sensitive, reliable in different settings around the world. We have developed a simple, rapid (<30 ​min), and inexpensive test for SARS-CoV-2 that is based on combination of isothermal reverse transcription recombinase polymerase amplification (RT-RPA) using modified primers and visual detection with paper-based microfluidics. Our test (CoRapID) is specific for SARS-CoV-2 (alpha to omicron variants) and does not detect other coronaviruses and pathogens by in silico and in vitro analysis. A two-step test protocol was developed with stable lyophilized reagents that reduces handling by using portable and disposable components (droppers, microapplicators/swabs, paper-strips). After optimization of assay components and conditions, we have achieved a limit of detection (LoD) of 1 copy/reaction by adding a blocking primer to the lateral flow assay. Using a set of 138 clinical samples, a sensitivity of 88.1% (P ​< ​0.05, CI: 78.2–93.8%) and specificity of 93.9% (P ​< ​0.05, CI: 85.4–97.6%) was determined. The lack of need for instrumentation for our CoRapID makes it an ideal on-site primary screening tool for local hospitals, doctors’ offices, senior homes, workplaces, and in remote settings around the world that often do not have access to clinical laboratories.

对新出现的病原体进行快速现场诊断是早期识别感染者和防止在人群中进一步传播的关键。目前可用的分子诊断测试是基于仪器的,而快速抗体和抗原测试通常对症状前受试者的检测不够敏感。需要快速的护理点分子筛查测试,这种测试可以很容易地适应新出现的病原体,并且在世界各地的不同环境中具有选择性、敏感性和可靠性。我们开发了一种简单、快速(<;30​min),以及基于使用改良引物的等温逆转录重组酶聚合酶扩增(RT-RPA)和纸基微流体的视觉检测的组合的廉价的SARS-CoV-2检测。我们的测试(CoRapID)对严重急性呼吸系统综合征冠状病毒2型(α-奥密克戎变异株)具有特异性,通过计算机和体外分析无法检测到其他冠状病毒和病原体。使用稳定的冻干试剂制定了两步测试方案,通过使用便携式和一次性组件(滴管、微涂抹器/拭子、纸条)减少了操作。在优化测定组分和条件后,我们通过向侧流测定中添加阻断引物,实现了1拷贝/反应的检测极限(LoD)。使用138个临床样本,敏感性为88.1%(P​<;​0.05,CI:78.2–93.8%),特异性为93.9%(P​<;​0.05,CI:85.4–97.6%)。我们的CoRapID不需要仪器,这使它成为当地医院、医生办公室、养老院、工作场所以及世界各地通常无法进入临床实验室的偏远地区的理想现场初级筛查工具。
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引用次数: 1
Parvovirus B19: Insights and implication for pathogenesis, prevention and therapy 细小病毒B19:发病机制、预防和治疗的见解和意义
Pub Date : 2023-01-01 DOI: 10.1016/j.amolm.2023.100007
K. Zakrzewska , R. Arvia , G. Bua , F. Margheri , G. Gallinella

Parvovirus B19 (B19V) is a small ssDNA non-enveloped virus, member of Parvoviridae family. The infection is widely diffused and is responsible for a broad range of clinical manifestations including fifth disease in children, transient aplastic crisis in patients with haematological disorders, non-immune hydrops fetalis in pregnant women, persistent anaemia in immunocompromised patients, arthropathy and inflammation of various other tissues. B19V infects and replicates in erythroid progenitor cells (EPCs) in the bone marrow. The depletion of infected EPCs represents the pathogenetic mechanisms of some haematological B19V-associate diseases.

Following a primary infection, the virus can establish lifelong persistence in several tissues. Currently, the pathological potential of persistent virus on the cellular signalling pathways remains unclear. In non-erythroid tissues, the infection is usually, abortive, and the virus seems to exert its pathological role through indirect mechanisms, such as induction of inflammatory and autoimmune processes, or through virus-induced apoptosis mediated by viral proteins. In addition to the diseases for which the etiological role of B19V has been fully demonstrated, there are several clinical conditions, including autoimmune diseases, that are presumably, but not certainly, associated with B19V infection.

In this review, we describe recent findings that may give us new insight into the pathogenic role of B19V in systemic sclerosis, an autoimmune disease of unknown multifactorial aetiology. Furthermore, we describe the latest findings on the intrauterine B19V infections. Moreover, since there are some ongoing interesting studies focused on vaccine development and antiviral drug discovery for the prevention and treatment of parvovirus B19 infection we described some advances in this field of research.

细小病毒B19 (B19V)是细小病毒科的一种小型非包膜病毒。这种感染广泛扩散,导致多种临床表现,包括儿童的第五种疾病、血液学疾病患者的一过性再生危象、孕妇的非免疫性水肿胎儿、免疫功能低下患者的持续贫血、关节病和各种其他组织的炎症。B19V在骨髓中的红系祖细胞(EPCs)中感染和复制。感染EPCs的耗竭代表了一些血液学b19v相关疾病的发病机制。初次感染后,病毒可在几个组织中建立终身持久性。目前,持续病毒在细胞信号通路上的病理潜力尚不清楚。在非红系组织中,感染通常是流产的,病毒似乎通过间接机制发挥其病理作用,如诱导炎症和自身免疫过程,或通过病毒蛋白介导的病毒诱导的细胞凋亡。除了B19V的病原学作用已得到充分证实的疾病外,还有一些临床疾病,包括自身免疫性疾病,可能与B19V感染有关,但不确定。在这篇综述中,我们描述了最近的研究结果,这些发现可能使我们对B19V在系统性硬化症(一种未知多因素病因的自身免疫性疾病)中的致病作用有了新的认识。此外,我们描述了宫内B19V感染的最新发现。此外,由于有一些正在进行的有趣的研究集中在疫苗开发和抗病毒药物的发现,以预防和治疗细小病毒B19感染,我们描述了这一研究领域的一些进展。
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引用次数: 1
TTV and other anelloviruses: The astonishingly wide spread of a viral infection TTV和其他类病毒:一种病毒感染的惊人的广泛传播
Pub Date : 2023-01-01 DOI: 10.1016/j.amolm.2023.100006
Pietro Giorgio Spezia , Daniele Focosi , Andreina Baj , Federica Novazzi , Francesca Drago Ferrante , Fabrizio Carletti , Claudia Minosse , Giulia Matusali , Fabrizio Maggi

The broad family of viruses known as anelloviruses (AV) infects both humans and numerous animal species. They have a tiny, covalently closed single-stranded DNA genome and the astonishing capacity to infect a very high percentage of healthy and ill people with chronic infections that could last a lifetime. AV, and particularly the prototype Torquetenovirus, have established a successful interaction with the host's immune system and the rate at which they replicate is a gauge to measure overall immune function, even though many aspects of their life cycle and pathogenesis are still poorly understood.

被称为anelloviruses(AV)的广泛病毒家族感染人类和许多动物物种。它们有一个微小的、共价封闭的单链DNA基因组,具有惊人的能力,可以感染高比例的健康人和病人,使其感染可能持续一生的慢性感染。AV,尤其是Torquetenovirus的原型,已经与宿主的免疫系统建立了成功的相互作用,它们的复制速度是衡量整体免疫功能的一个指标,尽管它们生命周期和发病机制的许多方面仍知之甚少。
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引用次数: 3
Extracellular vesicles engagement during respiratory viruses infection 呼吸道病毒感染时细胞外囊泡接合
Pub Date : 2023-01-01 DOI: 10.1016/j.amolm.2023.100004
Maria Alfreda Stincarelli , Rosaria Arvia , Simone Giannecchini

Respiratory viruses infection is a worldwide human concern annually. The main viral respiratory diseases are caused by a variety of viruses sharing similar threats and affecting the respiratory system. Among all, influenza viruses, respiratory syncytial virus, parainfluenza viruses, respiratory adenoviruses, rhinoviruses, human bocaviruses, human metapneumovirus and coronaviruses are the main common respiratory viruses affecting human population. The recent coronavirus disease 19 pandemic has revealed critical knowledge gaps required to update in the transmission and pathological induced pathways for respiratory viruses. To date, several evidences suggest that human viruses can hijack extracellular vesicles (EVs) to deliver proteins, mRNAs, microRNAs and whole viral particles during viral life cycle in the host. Thus, several investigations have reported that also respiratory viruses use EVs to deliver viral nucleic acid and proteins, even including the potentiality of carrying whole viral particle. This evidence demonstrates the ability of the EVs produced in infected cells to deliver respiratory viral components to uninfected cells, positively or negatively counteracting new viral infection. Additionally, EVs derived from biological fluids of clinical samples may increase the risk to induce severe respiratory viruses-associated diseases in site far from the respiratory tract and for prolonged time. Here, it has been reviewed the advantages of the respiratory viruses EVs interaction regarding their ability to enhance viral infection, to evade antiviral response, to regulate virus-immune response and to mediate diseases. All these data confirm a potential role of the association between EVs and respiratory viruses infection. This suggests that further studies to define the implication of this interaction in viral life cycle in human population are needed.

呼吸道病毒感染每年都是全世界人类关注的问题。主要的病毒性呼吸道疾病是由多种病毒引起的,它们具有相似的威胁并影响呼吸系统。其中,流感病毒、呼吸道合胞病毒、副流感病毒、呼吸道腺病毒、鼻病毒、人牛瘟病毒、人偏肺病毒和冠状病毒是影响人群的主要常见呼吸道病毒。最近的冠状病毒病19大流行揭示了需要更新呼吸道病毒传播和病理诱导途径的关键知识空白。迄今为止,一些证据表明,人类病毒可以劫持细胞外囊泡(EVs)在宿主体内的病毒生命周期中传递蛋白质、mrna、microrna和整个病毒颗粒。因此,一些研究报道呼吸道病毒也使用ev来传递病毒核酸和蛋白质,甚至包括携带整个病毒颗粒的可能性。这一证据表明,在感染细胞中产生的ev能够将呼吸道病毒成分传递给未感染的细胞,从而积极或消极地抵消新的病毒感染。此外,从临床样本的生物体液中提取的ev可能会增加在远离呼吸道的地方诱发严重呼吸道病毒相关疾病的风险,并可能持续较长时间。本文综述了呼吸道病毒与ev相互作用在增强病毒感染、逃避抗病毒反应、调节病毒免疫反应和介导疾病等方面的优势。所有这些数据都证实了ev与呼吸道病毒感染之间的潜在关联。这表明需要进一步的研究来确定这种相互作用在人类病毒生命周期中的含义。
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引用次数: 0
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Aspects of molecular medicine
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