Virology最新文献_第10页

Isolation, propagation, and characterization of a G9P[4] human rotavirus strain 543765 in Iran 伊朗g9p[4]人轮状病毒株543765的分离、繁殖和鉴定

IF 2.4 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-09-17 DOI: 10.1016/j.virol.2025.110695

Atefeh Kachooei , Reza Aramideh-Khouy , Mehrnaz Hosseini-Tehrani , Mahtab Mir-Hosseinian , Zahra Habib , Maryam Kazemi-Aghdam , Somayeh Jalivand , Tayebeh Latifi , Angila Ataei-Pirkooh , Zabihollah Shoja

Group A rotavirus (RVA) is a leading etiological agent of diarrheal diseases in children less than 5 years of age. While live attenuated RV vaccines have demonstrated high efficacy in high-income countries (HICs), their performance is substantially reduced in low- and middle-income countries (LMICs). Despite this disparity, the development and evaluation of live attenuated RVA strains remain a central objective in RV vaccine research. In this study, a human RVA strain, designated 543765, was successfully isolated from a stool sample using MA104 cell culture. The isolate was characterized through observation of cytopathic effects (CPE), polyacrylamide gel electrophoresis (PAGE), reverse transcription-polymerase chain reaction (RT-PCR), hemagglutination assay, transmission electron microscopy (TEM), and complete genome sequencing. Genotypic analysis revealed the following constellation: G9-P[4]-I1 (Lineage IV/II recombinant)-R1-C1-M1-A1-N1-T1-E1-H1. These findings suggest that strain 543765 exhibits stable structural and replication properties, achieving titers of up to 10⁸ TCID₅₀/mL in MA104 cells. Given its genetic profile and in vitro growth characteristics, strain 543765 holds promise as a candidate for development into a monovalent vaccine capable of inducing both homotypic and heterotypic immune protection against G9P[4] and other RVA genotypes. However, further investigation is warranted to evaluate whether serial passages in cell culture have resulted in attenuation, a determination that requires validation through clinical studies.

A组轮状病毒（RVA）是5岁以下儿童腹泻病的主要病原。虽然RV减毒活疫苗在高收入国家（HICs）显示出很高的效力，但在低收入和中等收入国家（LMICs），其效力却大大降低。尽管存在这种差异，开发和评估RVA减毒活株仍然是RV疫苗研究的中心目标。在本研究中，使用MA104细胞培养成功地从粪便样本中分离出了一株命名为543765的人RVA菌株。通过观察细胞病变效应（CPE）、聚丙烯酰胺凝胶电泳（PAGE）、逆转录聚合酶链反应（RT-PCR）、血凝试验、透射电镜（TEM）和全基因组测序对分离物进行了鉴定。基因型分析显示：G9-P[4]-I1（谱系IV/II重组）-R1-C1-M1-A1-N1-T1-E1-H1。这些结果表明，菌株543765具有稳定的结构和复制特性，在MA104细胞中滴度高达108 TCID50/mL。鉴于其遗传特征和体外生长特性，菌株543765有望开发成一种单价疫苗，能够诱导针对G9P[4]和其他RVA基因型的同型和异型免疫保护。然而，需要进一步的研究来评估细胞培养中的连续传代是否会导致衰减，这需要通过临床研究来验证。

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引用次数: 0

The VP7 protein of the African horse sickness virus core particle facilitates binding to Culicoides sonorensis cells in an RGD-independent manner 非洲马病病毒核心颗粒的VP7蛋白以不依赖rgd的方式促进与索诺库蠓细胞的结合。

IF 2.4 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-09-15 DOI: 10.1016/j.virol.2025.110694

Ariel Renée Monique Buyens, Vida van Staden, Jacques Theron

African horse sickness, caused by African horse sickness virus (AHSV) that is transmitted by midges of the Culicoides genus, leads to rapid mortality among horses. Proteases in the saliva of Culicoides midges cleave the VP2 outer capsid protein, resulting in infectious sub-virus particles that have increased infectivity for the Culicoides vector insect and Culicoides-derived cells (KC cells). The AHSV VP7 protein has an arginine-glycine-aspartate (RGD) motif, but the functional relevance of this protein and motif in facilitating binding to insect cells is unknown. To investigate, core-like particles (CLPs) were produced using the baculovirus expression system through the co-expression of VP3 and sVP7, which is a soluble version of the AHSV-4 VP7 protein. Insect cell binding assays indicated that the CLPs bind to KC cells, suggesting a role for VP7 in this interaction. Subsequently, recombinant baculoviruses expressing mutant sVP7 proteins were synthesized, in which the RGD motif was either deleted or mutated. All RGD-mutated sVP7 proteins, except for the deletion of the RGD motif, formed trimers and, when co-expressed with VP3, assembled into CLPs that retained the ability to bind to insect cells. These findings indicate that VP7 facilitates the binding of CLPs to insect cells through an RGD-independent mechanism.

非洲马病是由库蠓属的蠓传播的非洲马病病毒（AHSV）引起的，导致马迅速死亡。库蠓唾液中的蛋白酶可裂解VP2外衣壳蛋白，产生感染性亚病毒颗粒，增强库蠓媒介昆虫和库蠓衍生细胞（KC细胞）的传染性。AHSV VP7蛋白具有精氨酸-甘氨酸-天冬氨酸（RGD）基序，但该蛋白和基序在促进与昆虫细胞结合方面的功能相关性尚不清楚。为了进行研究，利用杆状病毒表达系统通过VP3和sVP7 （AHSV-4 VP7蛋白的可溶性版本）的共表达产生了核样颗粒（CLPs）。昆虫细胞结合实验表明，CLPs与KC细胞结合，表明VP7在这种相互作用中起作用。随后，合成了表达突变sVP7蛋白的重组杆状病毒，其中RGD基序被删除或突变。除了RGD基序缺失外，所有RGD突变的sVP7蛋白都形成三聚体，当与VP3共表达时，组装成CLPs，保留了与昆虫细胞结合的能力。这些发现表明VP7通过一种不依赖于rgd的机制促进CLPs与昆虫细胞的结合。

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引用次数: 0

A synonymous mutation in the preS2 region enhances production of infectious hepatitis B virus preS2区域的同义突变增强了传染性乙型肝炎病毒的产生。

IF 2.4 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-09-12 DOI: 10.1016/j.virol.2025.110692

Asako Murayama , Norie Yamada , Masaaki Toyama , Hussein Hassan Aly , Hironori Nishitsuji , Kunitada Shimotohno , Masanori Isogawa , Takanobu Kato

Background

Hepatitis B virus (HBV) is classified into at least nine genotypes based on sequence heterogeneity. Clinical and virological characteristics vary among these genotypes, and differences have also been reported among strains within the same genotype. In this study, we aimed to clarify the strain-specific characteristics of patient-derived genotype C (GT-C) strains and identify a synonymous mutation responsible for these characteristics, along with the underlying mechanisms.

Materials and methods

HBV molecular clones were constructed from sequences obtained from two chronic hepatitis patients infected with GT-C. To evaluate HBsAg production and infectivity, these molecular clones were transfected into cell cultures, and the characteristics of the generated viruses were assessed. The HBV reporter virus was used to confirm these characteristics and determine the responsible regions. mRNA quantification and mRNA transfection experiments were performed to elucidate the mechanisms underlying high HBsAg production and enhanced infectivity.

Results

HBsAg production and infectivity were analyzed in two GT-C strains, GT-C1 and GT-C2. GT-C2 exhibited higher HBsAg production than GT-C1 did, whereas GT-C1 showed greater infectivity. Analysis of chimeric and mutated strains revealed that a synonymous mutation, a3210g, in the preS2 region was responsible for the high HBsAg production of GT-C2. Introducing this mutation into the GT-C1 strain led to increased HBsAg production due to increased HBsAg translation efficiency and further enhanced infectivity.

Conclusions

This HBV infection system with both high HBsAg production and high infectivity provides a valuable tool for studying HBV infection and propagation in cell culture and for developing antiviral strategies for HBV infection.

背景：乙型肝炎病毒（HBV）根据序列异质性被分为至少9种基因型。这些基因型的临床和病毒学特征各不相同，并且在同一基因型的菌株之间也有差异的报道。在这项研究中，我们旨在阐明患者源性基因型C （GT-C）菌株的菌株特异性特征，并确定导致这些特征的同义突变，以及潜在的机制。材料与方法：从2例慢性肝炎患者的GT-C感染序列中构建HBV分子克隆。为了评估HBsAg的产生和传染性，将这些分子克隆转染到细胞培养物中，并评估所产生病毒的特性。使用HBV报告病毒来确认这些特征并确定负责区域。通过mRNA定量和mRNA转染实验来阐明高HBsAg产生和增强传染性的机制。结果：分析了GT-C1和GT-C2两株GT-C的HBsAg生成和感染性。GT-C2表现出比GT-C1更高的HBsAg生成，而GT-C1表现出更大的传染性。对嵌合和突变菌株的分析表明，在preS2区域的a3210g同源突变是GT-C2高HBsAg产生的原因。将这一突变引入GT-C1菌株，由于HBsAg翻译效率的提高和进一步增强的传染性，导致HBsAg产量增加。结论：该HBV感染系统具有高HBsAg生成和高传染性，为研究HBV在细胞培养中的感染和繁殖以及制定HBV感染的抗病毒策略提供了有价值的工具。

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引用次数: 0

Human cytomegalovirus interleukin 10 (cmvIL-10) orchestrates crosstalk between the cellular IL-10 receptor and CXCR4 人巨细胞病毒白细胞介素10 （cmvIL-10）协调细胞IL-10受体和CXCR4之间的串扰

IF 2.4 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-09-12 DOI: 10.1016/j.virol.2025.110693

Kiran H. Satani , Juliet V. Spencer

Human cytomegalovirus (HCMV) is a widespread beta-herpesvirus that establishes lifelong latent infection in host cells. The HCMV UL111A gene encodes cmvIL-10, a viral homolog of the anti-inflammatory cytokine IL-10 (hIL-10). Like hIL-10, cmvIL-10 binds to the human cellular IL-10 receptor (IL-10R) and activates the JAK/STAT3 pathway, promoting transcription of immunosuppressive genes. Notably, cmvIL-10 binds to the IL-10R with higher affinity than hIL-10 and retains most of its functions, including enhancing CXCR4 signaling triggered by its primary ligand, CXCL12. The CXL12/CXCR4 axis regulates key cellular processes such as gene expression, cell migration, proliferation, and apoptosis. CXCR4 also interacts with alternative ligands including macrophage inhibitory factor (MIF), trefoil factor 2 protein (TFF2), and extracellular ubiquitin (UB). Here, we show that cmvIL-10 enhances CXCR4-mediated cell proliferation and migration induced by MIF and TFF2 but not by UB. We also found that hIL-10 enhances MIF-mediated cell proliferation yet has no effect on TFF2-mediated cell proliferation. The CXCR4 antagonist AMD3100 effectively blocks cell migration driven by TFF2, while only partially inhibiting MIF-mediated cell migration. These results suggest that cmvIL-10 promotes crosstalk between IL-10R and CXCR4, underscoring its role in modulating immune responses during HCMV infection. Our findings provide insight into how cmvIL-10 contributes to immune evasion and pathogenesis in HCMV.

人巨细胞病毒（HCMV）是一种广泛存在的乙型疱疹病毒，可在宿主细胞中建立终身潜伏感染。HCMV UL111A基因编码cmvIL-10，这是一种抗炎细胞因子IL-10 （hIL-10）的病毒同源物。与hIL-10一样，cmvIL-10与人细胞IL-10受体（IL-10R）结合，激活JAK/STAT3通路，促进免疫抑制基因的转录。值得注意的是，cmvIL-10以比il -10更高的亲和力与IL-10R结合，并保留其大部分功能，包括增强其主要配体CXCL12触发的CXCR4信号转导。CXL12/CXCR4轴调控关键的细胞过程，如基因表达、细胞迁移、增殖和凋亡。CXCR4还与其他配体相互作用，包括巨噬细胞抑制因子（MIF）、三叶因子2蛋白（TFF2）和细胞外泛素（UB）。在这里，我们发现cmvIL-10可以增强MIF和TFF2诱导的cxcr4介导的细胞增殖和迁移，而UB则不能。我们还发现，hIL-10增强了mif介导的细胞增殖，而对tff2介导的细胞增殖没有影响。CXCR4拮抗剂AMD3100有效阻断TFF2驱动的细胞迁移，而仅部分抑制mif介导的细胞迁移。这些结果表明cmvIL-10促进IL-10R和CXCR4之间的串扰，强调其在HCMV感染期间调节免疫应答中的作用。我们的研究结果为cmvIL-10如何参与HCMV的免疫逃避和发病机制提供了见解。

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引用次数: 0

Regulatory T cell infiltration precedes early events associated with persistent infectious bronchitis virus (IBV) infection in the cecal tonsils of chickens 调节性T细胞浸润先于鸡盲肠扁桃体持续性传染性支气管炎病毒（IBV）感染相关的早期事件。

IF 2.4 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-09-12 DOI: 10.1016/j.virol.2025.110691

Sufna M. Suhail, Ishara M. Isham, Muhammad Farooq, Muhammad Azhar, Ahmed Ali, Motamed E. Mahmoud, Awais Ghaffar, Heshanthi Herath Mudiyansalage, Susan C. Cork, Ashish Gupta, Mohamed Faizal Abdul-Careem

Infectious bronchitis (IB) is a contagious respiratory disease in chickens caused by infectious bronchitis virus (IBV). While initially affecting the respiratory tract, IBV has evolved to infect other body systems. Notably, the Delmarva (DMV)/1639 genotype of IBV has been shown to persist in the cecal tonsils (CT), potentially facilitating viral transmission to naïve birds. This study aimed to uncover the early immunological mechanisms leading up to IBV persistence in the CT, compared to the spleen and to evaluate the persistence patterns among different genotypes of IBV.

An animal experiment was conducted using three IBV genotypes, with samples collected at 3, 8, 10, and 14 days post-infection (dpi). Across all IBV infected groups, viral genome loads were significantly higher in the CT than that in the spleen. Recruitment of B cells and cluster of differentiation (CD)8⁺ T cells, crucial for viral clearance, was significantly lower in the CT. Regulatory T (Treg) cells—which suppress immune responses via interleukin (IL)-10 and transforming growth factor (TGF)-β—were expected to peak early, their abundance in the CT was significantly higher only at the later time points. Among all genotypes, DMV/1639 exhibited increased potential for persistence in the CT associated with higher levels of Treg cells and viral genome load at 14 dpi.

These findings suggest that delayed Treg cell infiltration, reduced effector cell recruitment, and a suppressive cytokine environment may precede to IBV persistence in the CT. Further studies are needed to explore the potential role of anti-inflammatory cytokines and other immunological factors in IBV persistence in the CT.

传染性支气管炎（IB）是由传染性支气管炎病毒（IBV）引起的鸡传染性呼吸道疾病。虽然最初影响呼吸道，但IBV已经进化到感染其他身体系统。值得注意的是，IBV的Delmarva (DMV)/1639基因型已被证明在盲肠扁桃体（CT）中持续存在，可能促进病毒向naïve鸟类传播。本研究旨在揭示导致IBV在CT和脾脏中持续存在的早期免疫学机制，并评估不同基因型IBV的持续模式。采用三种IBV基因型进行动物实验，分别于感染后3、8、10和14天采集样本。在所有IBV感染组中，CT中的病毒基因组载量明显高于脾脏中的病毒基因组载量。在CT中，对病毒清除至关重要的B细胞和8+ T细胞的募集明显减少。通过白细胞介素(IL)-10和转化生长因子(TGF)-β抑制免疫反应的调节性T （Treg）细胞预计会在早期达到峰值，其在CT中的丰度仅在较晚的时间点才显着升高。在所有基因型中，DMV/1639在CT中表现出更高的持久性潜力，这与14 dpi时更高水平的Treg细胞和病毒基因组载量有关。这些结果表明，延迟Treg细胞浸润、减少效应细胞募集和抑制细胞因子环境可能是IBV在CT中持续存在的原因。抗炎细胞因子和其他免疫因子在IBV CT持续性中的潜在作用有待进一步研究。

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引用次数: 0

Comparative coat protein annotation of two biologically distinct strains of cucumber mosaic virus in chili 辣椒中黄瓜花叶病毒两种不同菌株外壳蛋白的比较注释

IF 2.4 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-09-12 DOI: 10.1016/j.virol.2025.110690

Vinodhini Jeevarathinam , Rajendran Lingan , Jeya Sundara Sharmila , Karthikeyan Gandhi

Cucumber mosaic virus (CMV) is a globally prevalent plant virus with a broad host range, significantly affecting chili crops. From a diagnostic survey, 49 symptomatic chili plants were collected and CMV was confirmed in 23 samples. Molecular indexing revealed a predominance of mosaic-inducing strains (18 samples) along with distinct chlorosis inducing strains (5 samples) of CMV in the chili eco system of Tamil Nadu. Representative strains were mechanically sap inoculated onto chili host plants and produced similar symptoms as observed under field conditions. Coat protein (CP) gene analysis revealed 657 nucleotides encoding for 218 amino acids with a homology of 97.65 %–98.92 % with Indian chili isolates. Sequence diversity analysis indicated the higher synonymous substitutions in chlorosis-inducing strains, suggesting greater nucleotide variability. Comparative amino acid analysis indicated conserved residues in chlorosis-inducing strains such as Ser¹²⁹, Thr¹³⁷ and Thr¹⁶² and mosaic-inducing strains such as Pro¹²⁹, Ser¹³⁷ and Ala¹⁶². Furthermore, CP annotation performed by predicting secondary structures showed conformational modification in the loop structure of protein. To validate the symptomatology, in silico protein-protein docking was carried out to explore the possible interaction between chloroplast ferredoxin proteins of chili based on previous studies. Functional annotation substantiated that chlorotic symptom expression may be due to amino acid variation of Ser¹²⁹ and aids compatible interaction between chloroplast ferredoxin proteins of chili. These findings provide novel insights into the molecular basis of strain-specific symptom development and highlight the need for targeted diagnostics of emerging CMV variants in chili cultivation.

黄瓜花叶病毒（CMV）是一种全球流行的植物病毒，寄主范围广，严重影响辣椒作物。从诊断调查中，收集了49株有症状的辣椒，其中23株被确认为巨细胞病毒。分子指数分析显示，泰米尔纳德邦辣椒生态系统中CMV的嵌合诱导菌株（18个样本）占主导地位，而致绿诱导菌株（5个样本）则不同。将有代表性的菌株机械接种到辣椒寄主植株上，产生的症状与田间条件下观察到的相似。外皮蛋白（CP）基因与印度辣椒的同源性为97.65% ~ 98.92%，共657个核苷酸编码218个氨基酸。序列多样性分析表明，致绿变菌株具有较高的同义替换，表明核苷酸具有较大的变异性。氨基酸比较分析表明，在褪绿诱导菌株Ser129、Thr137、Thr162和嵌合诱导菌株Pro129、Ser137、Ala162中存在保守残基。此外，通过预测二级结构进行的CP注释显示了蛋白质环结构的构象改变。为了验证症状学，在前人研究的基础上，进行硅蛋白-蛋白对接，探讨辣椒叶绿体铁氧化还蛋白之间可能的相互作用。功能注释证实了变色症状的表达可能是由于Ser129的氨基酸变异，并有助于辣椒叶绿体铁氧还蛋白之间的相容相互作用。这些发现为菌株特异性症状发展的分子基础提供了新的见解，并强调了在辣椒栽培中对新出现的巨细胞病毒变异进行靶向诊断的必要性。

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引用次数: 0

Breast milk lacking anti-human immunodeficiency virus activity promotes exocytosis of HIV from the mother's mammary epithelium and transcytosis of virus via the infant's tonsil and intestinal epithelial cells 缺乏抗人类免疫缺陷病毒活性的母乳促进母亲乳腺上皮细胞的HIV胞吐和病毒通过婴儿的扁桃体和肠上皮细胞的胞吞

IF 2.4 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-09-11 DOI: 10.1016/j.virol.2025.110689

Nicole T. Padilla , Xiaodan Cai , Rossana Herrera , Kristina Rosbe , Sharof M. Tugizov

Although mammary epithelial cells (MECs) and breast milk facilitate mother-to-child transmission (MTCT) of human immunodeficiency virus (HIV-1), the mechanisms underlying these observations remain poorly understood. In this work, we explored the molecular mechanisms associated with HIV-1 transcytosis through MECs and the role of breast milk in promoting viral transmigration through infant tonsils and intestinal epithelia. Our findings revealed that transfer of cell-free HIV-1 from the maternal circulation into breast milk is mediated by its basolateral-to-apical transcytosis through polarized MECs that function as a blood-milk barrier. While breast milk samples from 76 % of the HIV-negative donors contained factors that inactivated the virus by disrupting viral envelope glycoprotein gp120, thereby preventing MTCT, the remaining breast milk samples that were incapable of inactivating HIV-1 nonetheless induced viral exocytosis from the apical surface of MECs. Interestingly, these otherwise inactive breast milk samples also promoted a substantial increase in viral internalization, transcytosis, and exocytosis from infant tonsils and gut epithelial cells, ultimately leading to infection of virus-susceptible subepithelial cells. We determined that elevated calcium concentrations in breast milk play an important role in promoting HIV-1 entry, transcytosis, and exocytosis from infant tonsil and gut epithelial cells via activation of protein kinase C and calcium sensor synaptotagmin 7. Collectively, these findings suggest that breast milk samples from different sources may either promote or prevent HIV MTCT by several different mechanisms. Further investigation of this phenomenon may ultimately improve our understanding of the molecular pathogenesis of HIV MTCT and suggest new strategies for its prevention.

尽管乳腺上皮细胞（MECs）和母乳促进了人类免疫缺陷病毒（HIV-1）的母婴传播（MTCT），但这些观察结果背后的机制仍然知之甚少。在这项工作中，我们探讨了通过mec与HIV-1胞吞作用相关的分子机制，以及母乳在促进病毒通过婴儿扁桃体和肠上皮细胞转运中的作用。我们的研究结果表明，无细胞HIV-1从母体循环转移到母乳中是通过极化mec从基底外侧到根尖的胞饮作用介导的，mec作为血乳屏障发挥作用。虽然76%的hiv阴性供者的母乳样本含有通过破坏病毒包膜糖蛋白gp120来灭活病毒的因子，从而阻止MTCT，但其余不能灭活HIV-1的母乳样本仍然诱导了mec顶端表面的病毒胞外分泌。有趣的是，这些原本不活跃的母乳样本也促进了婴儿扁桃体和肠道上皮细胞的病毒内化、胞吞和胞吐的大幅增加，最终导致病毒易感的上皮下细胞感染。我们确定母乳中钙浓度升高通过激活蛋白激酶C和钙传感器突触塔蛋白7，在促进婴儿扁桃体和肠道上皮细胞的HIV-1进入、胞吞和胞外分泌中起重要作用。总的来说，这些发现表明，来自不同来源的母乳样本可能通过几种不同的机制促进或预防艾滋病毒母婴传播。对这一现象的进一步研究可能最终提高我们对HIV MTCT分子发病机制的理解，并为其预防提供新的策略。

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引用次数: 0

Generation and application of novel DNA aptamers targeted to the nucleocapsid protein of the SARS-CoV-2 针对SARS-CoV-2核衣壳蛋白的新型DNA适体的生成及应用

IF 2.4 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-09-11 DOI: 10.1016/j.virol.2025.110688

Xinyue Zhang , Lingfeng Guan , Hanchuan Wang , Jiaxin Liu , Xinyan Wei , Zhenghong Xu , Shaowen Wang , Qiwei Qin

Coronavirus disease 2019 (COVID-19) is a respiratory disease caused by SARS-CoV-2, with common symptoms of fever, cough, and dyspnea. SARS-CoV-2 transmission with high prevalence includes aerosols, respiratory droplets, and fecal-oral routes. To date, COVID-19 has infected more than 7.7 million people, seriously threatening people around the world. Therefore, rapid diagnosis of COVID-19 is crucial for epidemic prevention and control. In this study, two novel DNA aptamers (apt1 and apt2) targeting the nucleocapsid protein of the SARS-CoV-2 (CoV2-N) were screened through selective evolution of ligands by exponential enrichment (SELEX). Subsequently, we devised a simple and sensitive lateral flow biosensor (LFB) for quick detection of the CoV2-N. Through secondary structure and minimum free energy predictions, we found that both apt1 and apt2 form stable stem-loop structures, with free energies of −16.01 and −14.04 kJ/mol, respectively. Moreover, they showed high binding affinities to CoV2-N, with calculated binding affinities (K_d) of 18.9 nM for apt1 and 55.7 nM for apt2. The aptamer-based LFB features simple operation, with a detection limit of 500 ng/mL for proteins and visually interpretable results within 5 min. Together, we obtained aptamers that specifically recognized CoV2-N and established a new detection method with high specificity and sensitivity.

冠状病毒病2019 （COVID-19）是一种由SARS-CoV-2引起的呼吸道疾病，常见症状为发烧、咳嗽和呼吸困难。高流行率的SARS-CoV-2传播途径包括气溶胶、呼吸道飞沫和粪口途径。迄今为止，COVID-19已感染770多万人，严重威胁着世界各地的人民。因此，快速诊断COVID-19对于疫情防控至关重要。本研究通过指数富集（SELEX）配体的选择性进化，筛选了靶向SARS-CoV-2 （CoV2-N）核衣壳蛋白的两个新型DNA适体（apt1和apt2）。随后，我们设计了一种简单灵敏的横向流动生物传感器（LFB），用于快速检测CoV2-N。通过二级结构和最小自由能预测，我们发现apt1和apt2均形成稳定的茎环结构，自由能分别为- 16.01和- 14.04 kJ/mol。此外，它们对CoV2-N具有较高的结合亲和力，计算出apt1的结合亲和力（Kd）为18.9 nM， apt2为55.7 nM。基于适配体的LFB操作简单，对蛋白质的检出限为500 ng/mL，结果在5 min内可直观解释。我们共同获得了特异性识别CoV2-N的适配体，建立了一种特异性和灵敏度高的检测新方法。

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引用次数: 0

Editorial: 70th Anniversary of Virology 社论：病毒学成立70周年。

IF 2.4 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-09-10 DOI: 10.1016/j.virol.2025.110687

Jasmine Tomar (Editor-in-Chief)

引用次数: 0

Characterization of phage vB_EfS_C1 and investigation of antibacterial and antibiofilm properties against vancomycin-susceptible and vancomycin-resistant Enterococcus faecium 噬菌体vB_EfS_C1的鉴定及对万古霉素敏感和耐万古霉素肠球菌的抑菌和抗菌膜性能研究

IF 2.4 3区医学 Q3 VIROLOGY

Virology

Pub Date : 2025-09-06 DOI: 10.1016/j.virol.2025.110685

Mohamed El-Telbany , Keigo Yamamura , Norihiro Saito , Yoshimitsu Masuda , Takahisa Miyamoto , Ken-ichi Honjoh

E. faecium is listed among the ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter), which are responsible for a significant proportion of global deaths. Vancomycin-resistant E. faecium (VREfm) is the leading species responsible for human infections, and the limited therapeutic options make VREfm a public health problem. This study aimed to isolate and characterize a novel phage against E. faecium and evaluate its activity against vancomycin-sensitive E. faecium (VSEfm) and VREfm. We isolated a phage, named vB_EfS_C1, from compost. The host range analysis showed that it produced plaques on 5 out of 18 enterococci strains and did not produce plaques on other species. Phage vB_EfS_C1 was infective at different temperatures (40–70 °C), and the titer was stable at different pH levels (6–9). The phage vB_EfS_C1 is 86,777 bp in length, grouping it with phages with large genomes infecting enterococci (>80 kbp). The killing curve revealed that phage vB_EfS_C1 had antibacterial activity with different MOIs (0.001–1) against the E. faecium host strain. The biofilm biomass of E. faecium FHC29 was reduced by 85 % after 6 and 24 h, and the biofilm biomass of E. faecium No. 982 (VREfm) by 50 % after 6 h and 25 % after 24 h with an MOI of 0.1. The viable counts of E. faecium FHC29 and VREfm in biofilms were decreased by 3 log and 1.83 log with an MOI of 1, respectively. In conclusion, phage vB_EfS_C1 is a lytic enterococcal phage displaying antibiofilm activity against VSEfm and VREfm.

粪肠杆菌被列为ESKAPE病原体（粪肠球菌、金黄色葡萄球菌、肺炎克雷伯菌、鲍曼不动杆菌、铜绿假单胞菌和肠杆菌）之一，这些病原体在全球死亡人数中占很大比例。万古霉素耐药粪肠杆菌（VREfm）是导致人类感染的主要物种，有限的治疗选择使VREfm成为一个公共卫生问题。本研究旨在分离和鉴定一种新的抗屎肠杆菌噬菌体，并评价其对万古霉素敏感屎肠杆菌（VSEfm）和VREfm的活性。我们从堆肥中分离出一个噬菌体，命名为vB_EfS_C1。宿主范围分析表明，它在18株肠球菌中的5株上产生斑块，而在其他种上不产生斑块。噬菌体vB_EfS_C1在不同温度（40 ~ 70℃）下均具有感染性，在不同pH值（6 ~ 9）下滴度稳定。噬菌体vB_EfS_C1全长86,777 bp，与感染肠球菌的大基因组噬菌体（>80 kbp）属一类。结果表明，噬菌体vB_EfS_C1对粪肠杆菌具有不同moi（0.001-1）的抑菌活性。粪肠杆菌FHC29的生物膜生物量在6 h和24 h后分别减少85%，粪肠杆菌982 （VREfm）的生物膜生物量在6 h和24 h后分别减少50%和25%，MOI为0.1。粪肠杆菌FHC29和VREfm在生物膜中的活菌数分别减少了3 log和1.83 log， MOI为1。综上所述，噬菌体vB_EfS_C1是一种具有抗VSEfm和VREfm活性的肠球菌噬菌体。

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