Journal of virological methods最新文献_第2页

Development of a native-antigen Western blot using ZZ-R 127–derived FMDV non-structural proteins for qualitative serology to inform DIVA workflows 使用zz - r127衍生的FMDV非结构蛋白进行定性血清学的原生抗原Western blot开发，为DIVA工作流程提供信息。

IF 1.6 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS

Journal of virological methods

Pub Date : 2026-05-01 Epub Date: 2026-02-06 DOI: 10.1016/j.jviromet.2026.115362

Fuat Özyörük , Ünal Parlak , Can Çokçalışkan , Santina Grazioli , Yılmaz Akça

This study describes the development of a Western blot (WB) assay employing native non-structural proteins (NSPs) from foot-and-mouth disease virus (FMDV)–infected ZZ-R 127 cell lysates to support confirmatory serodiagnosis within DIVA (differentiation of infected from vaccinated animals) frameworks. Native antigens were generated from six FMDV isolates representing serotypes A, O, and Asia1 and were separated by denaturing SDS–PAGE. Antigen identity was verified by co-migration with recombinant counterparts and detection with monoclonal antibodies (MAbs). Using one NSP-positive and one NSP-negative polyclonal bovine serum, the assay demonstrated consistent identification of 2C, 3D, and an ∼25.8 kDa 3AB-consistent precursor as the most reliable diagnostic targets. While MAb profiling confirmed the presence of 3C in the lysates, it showed no reactivity with the polyclonal serum, indicating limited utility for serology. Pixel-based quantification revealed predictable dilution-dependent signal behavior within gels (R² > 0.91 on log-log plots), though inter-gel variability was substantial, favoring qualitative over quantitative application. Absence of signals in mock-infected cell lysates and in the negative serum supported specificity. By preserving the native processing context of immunodominant NSPs, this platform is well-suited as a second-tier tool to adjudicate equivocal ELISA results in vaccinated populations.

本研究利用口蹄疫病毒（FMDV）感染ZZ-R 127细胞裂解物中的天然非结构蛋白（NSPs）开发了一种Western blot （WB）检测方法，以支持DIVA（感染动物与接种动物的区分）框架内的确证血清诊断。从代表A、O和Asia-1血清型的6株FMDV分离株中生成天然抗原，并通过变性SDS-PAGE分离。通过与重组对应物的共迁移和单克隆抗体（mab）检测来验证抗原的同一性。使用一份nsp阳性和一份nsp阴性的多克隆牛血清，该检测显示2C、3D和~25.8kDa 3ab一致的前体是最可靠的诊断靶标。虽然单抗分析证实了裂解物中存在3C，但它与多克隆血清无反应性，表明血清学用途有限。基于像素的量化揭示了凝胶内可预测的稀释依赖信号行为（对数对数图R2 > 0.91），尽管凝胶间的可变性很大，有利于定性而不是定量应用。在模拟感染的细胞裂解物和阴性血清中缺乏信号支持特异性。通过保留免疫显性NSPs的本地加工背景，该平台非常适合作为判定接种人群中模棱两可的ELISA结果的第二级工具。

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引用次数: 0

Inactivation of infectious virus samples through UV irradiation for downstream analysis 通过紫外线照射灭活传染性病毒样本用于下游分析。

IF 1.6 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS

Journal of virological methods

Pub Date : 2026-05-01 Epub Date: 2026-01-24 DOI: 10.1016/j.jviromet.2026.115354

Hugues Graf, Sébastien Faure, Maelle Mellano , Stéphanie Baron, Cynthia Chapel, Fanny Savoye, Eric Richier

Downstream analysis of infectious samples often needs to be performed outside a containment area, requiring prior inactivation before transfer. The inactivation of adenovirus, reovirus, and rabies virus by ultraviolet radiation at 254 nm (UV-C) was investigated with the objective of preserving the bacterial endotoxin test (BET) performance in irradiated samples. Virus samples containing endotoxins were exposed to UV-C doses ranging from 1 to 9 J/cm². A treatment of 5 J/cm² (2 × 2.5 J/cm²) applied to 1 mL volumes resulted in complete viral inactivation and acceptable endotoxin recovery. Samples containing cell culture media required 10-fold dilution prior to UV irradiation to overcome UV light attenuation and achieve complete virus inactivation. UV radiation had minimal impact on endotoxins, with recovery rates around 100 % compared to non-irradiated controls, regardless of UV dose and sample volume. The development of optimized UV-C irradiation conditions enabled the identification of a dose interval that ensures effective viral inactivation while maintaining BET specificity and applicability. The irradiation process is intended for industrial implementation on rabies vaccine samples, allowing testing outside containment areas and ensuring that biological samples are safely inactivated, without residual biological risk.

感染性样品的下游分析通常需要在控制区域之外进行，在转移前需要事先灭活。研究了254nm （UV-C）紫外线对腺病毒、呼肠孤病毒和狂犬病毒的灭活作用，目的是保持辐照样品的细菌内毒素试验（BET）性能。将含有内毒素的病毒样本暴露在1至9J/cm²的UV-C剂量下。5J/cm²（2 × 2.5J/cm²）1mL体积的处理导致病毒完全失活和可接受的内毒素恢复。含有细胞培养基的样品在紫外线照射前需要稀释10倍，以克服紫外线衰减并实现完全的病毒灭活。紫外线辐射对内毒素的影响最小，与未照射的对照组相比，无论紫外线剂量和样品量如何，其回收率约为100%。优化的UV-C照射条件的开发，使确定一个剂量间隔，确保有效的病毒灭活，同时保持BET的特异性和适用性。辐照过程旨在对狂犬病疫苗样本进行工业实施，允许在控制区域之外进行测试，并确保生物样本安全灭活，没有残留的生物风险。

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引用次数: 0

Prevalence and ribavirin-mediated elimination of Morchella importuna endornavirus 1 (MiEV1) in cultivated Morchella species in China 利巴韦林介导的羊肚菌内病毒1 （MiEV1）在中国养殖羊肚菌中的流行及消除

IF 1.6 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS

Journal of virological methods

Pub Date : 2026-05-01 Epub Date: 2026-01-12 DOI: 10.1016/j.jviromet.2026.115349

Zhifeng Zhang , Yingjun Zhang , Zengqiang Shang , Xiaoli Wang , Yankui Li , Zhaoyu Wang , Longbo Zhao , Ziyan Peng

Fungal viruses compromise the cultivation of Morchella. We surveyed five mycoviruses (Morchella importuna endornavirus 1 [MiEV1], Morchella importuna endornavirus 2 [MiEV2], Morchella importuna endornavirus 3 [MiEV3], Morchella esculenta fusarivirus 1 [MeFV1], and Morchella esculenta fusarivirus 2 [MeFV2]) in 24 cultivated strains (8 M. importuna, 5 M. septimelata, and 11 M. sextelata) collected across 11 provinces in China and evaluated candidate antiviral agents. Only MiEV1 was detected, with a high incidence of 79.2 % (19/24) across all three Morchella species. Among seven tested chemicals (ampicillin, azithromycin, kanamycin, rifampicin, roxithromycin, acyclovir hydrochloride, and ribavirin), only ribavirin significantly inhibited MiEV1 replication. Successive ribavirin treatments eliminated MiEV1 from a representative M. sextelata strain (M11), with the virus-free status remaining stable after multiple subcultures on drug-free medium. Elimination of MiEV1 also significantly enhanced mycelial growth rate and biomass, and led to enhanced mycelial pigmentation, suggesting a potential negative impact of MiEV1 on vegetative growth and pigmentation in M. sextelata. These findings demonstrate that MiEV1 is prevalent in cultivated Morchella and that ribavirin provides an effective strategy for generating stable virus-free strains, offering opportunities for mechanistic studies and industrial cultivation.

真菌病毒破坏羊肚菌的培养。我们对24株培养菌株（8 M.）进行了5种分枝病毒的检测，分别为：羊肚菌内膜病毒1型[MiEV1]、羊肚菌内膜病毒2型[MiEV2]、羊肚菌内膜病毒3型[MiEV3]、羊肚菌镰刀病毒1型[MeFV1]和羊肚菌镰刀病毒2型[MeFV2]。importuna 5 M。septimelata和11 M。在中国11个省份收集的六角星（sextelata），并评估候选抗病毒药物。仅检测到MiEV1，在所有3种羊肚菌中发病率高达79.2% %（19/24）。在7种化学物质（氨苄西林、阿奇霉素、卡那霉素、利福平、罗红霉素、盐酸阿昔洛韦和利巴韦林）中，只有利巴韦林能显著抑制MiEV1的复制。连续的利巴韦林治疗消除了具有代表性的六角分枝杆菌菌株（M11）中的MiEV1，在无药培养基上多次传代培养后，其无病毒状态保持稳定。消除MiEV1还能显著提高菌丝生长速率和生物量，并导致菌丝色素沉着增强，表明MiEV1对六爪草的营养生长和色素沉着有潜在的负面影响。这些发现表明，MiEV1在培养的羊肚菌中普遍存在，利巴韦林为产生稳定的无病毒菌株提供了有效的策略，为机理研究和工业化培养提供了机会。

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引用次数: 0

First report of laboratory-confirmed Zika virus infection in human from South Karnataka, India: A case study 印度南卡纳塔克邦首例经实验室确认的人感染寨卡病毒报告：一个案例研究。

IF 1.6 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS

Journal of virological methods

Pub Date : 2026-05-01 Epub Date: 2026-01-31 DOI: 10.1016/j.jviromet.2026.115357

Mallikarjun Koppad , Santhosh Kogaluru Shivakumaraswamy , Puneeth Thaduru Goolappa , Maithili Thotadur Mahabaleswara , Kasabi Gudadappa Siddappa , Premalatha Dindare Eswarappa , Siddesh Kaalavvala Channappa

Background

Zika virus is a member of flaviviruses group and is primarily transmitted by infected mosquito. Most of the Zika virus infections are subclinical or self limiting illness like dengue fever, but certain cases are manifested with Guillain-Barre syndrome in adults and microcephaly in neonates from infected mothers. Since 2016, India has considerably reported the Zika virus infection from different states (Gujarat, Tamil Nadu, Madhya Pradesh, Rajasthan, Kerala, Maharashtra and Uttar Pradesh). Similarly there was a case suspecting arboviral infection in Shivamogga city in Mid June, 2024 and found positive for Zika virus.

Method-

A serum sample of patient with hemorrhagic fever syndrome from private super speciality hospital suspecting for Zika virus infection was collected and sent to our laboratory for diagnosis of Zika virus. The sample was screened by real time reverse transcription polymerase chain reaction (RT-PCR) and confirmed by nucleotide sequencing. Further, the sample was examined for presence of anti-Zika IgM antibody. Finding- The real time RT-PCR result revealed the presence Zika virus RNA in the sample (CT value- 27) and it was further confirmed by sequencing the partial envelope (E) gene. Additionally, the subsequent serum sample was positive for anti-Zika IgM antibody. The phylogenetic analysis evidenced that Zika virus identified in this study belongs to the Asian lineage and was homogenous to the previously reported strains from India.

Interpretation

The patient without any travel history admitted for hemorrhagic fever was found positive with Zika virus. This is the first case of laboratory-confirmed Zika virus infection in human from Shivamogga, South Karnataka. It was further confirmed by phylogenetic analysis of envelope coding region and detecting anti-Zika virus antibody. However further studies have to be transacted to know the source of infection and systematic community based surveillance is need of the hour for early detection of Zika virus transmission.

背景：寨卡病毒是黄病毒科的一员，主要通过受感染的蚊子传播。大多数寨卡病毒感染是亚临床或自限性疾病，如登革热，但某些病例表现为成人格林-巴利综合征和受感染母亲所生的新生儿小头畸形。自2016年以来，印度不同邦（古吉拉特邦、泰米尔纳德邦、中央邦、拉贾斯坦邦、喀拉拉邦、马哈拉施特拉邦和北方邦）报告了大量寨卡病毒感染病例。同样，2024年6月中旬在Shivamogga市发生了一起疑似虫媒病毒感染的病例，并发现寨卡病毒呈阳性。方法：采集疑似寨卡病毒感染的私立超级专科医院出血热综合征患者血清标本，送实验室进行寨卡病毒诊断。通过实时反转录聚合酶链反应（RT-PCR）筛选样品，并进行核苷酸测序确认。此外，检测样本是否存在抗寨卡病毒IgM抗体。发现-实时RT-PCR结果显示样品中存在寨卡病毒RNA (CT值- 27)，并通过部分包膜(E)基因测序进一步证实。此外，随后的血清样本抗寨卡IgM抗体阳性。系统发育分析证明，本研究发现的寨卡病毒属于亚洲谱系，与先前报道的印度毒株同质。解释：无旅行史因出血热入院的患者被发现寨卡病毒阳性。这是来自南卡纳塔克邦Shivamogga的首例经实验室确认的人间寨卡病毒感染病例。通过包膜编码区系统发育分析和寨卡病毒抗体检测进一步证实。然而，必须进行进一步的研究，以了解感染源，并需要系统的社区监测，以便及早发现寨卡病毒传播。

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引用次数: 0

Animal models illuminate dengue immunopathogenesis to guide vaccine and therapeutic developments 动物模型阐明登革热的免疫发病机制，以指导疫苗和治疗的发展

IF 1.6 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS

Journal of virological methods

Pub Date : 2026-05-01 Epub Date: 2026-01-16 DOI: 10.1016/j.jviromet.2026.115350

Muhammad Nadir Shabbir , Thuy Linh Duong

Dengue virus (DENV) infection affects over 390 million people annually, and its severe forms, including dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS), occur as a result of antibody-dependent enhancement (ADE), T-cell dysregulation, and a cytokine storm. This review integrates the results of human immunopathology with those reported by animal models, consolidating the findings in a systematic manner to validate these mechanisms. Research on AG129 interferon receptor-deficient mice consistently demonstrates ADE-mediated viremia and vascular leakage. However, humanized NSG-BLT mice recapitulate the classic antigenic sin of human T cells after heterotypic challenge with DENV. Cynomolgus macaque models are physiologically relevant for testing tetravalent vaccines under near-human circulatory conditions, and diet-induced obese mice are used to phenotype the effects of metabolic comorbidities on IL-1β-driven immunopathology. Data using standardized methods, including Evans blue extravasation assays, multiplex cytokine panels, and endpoints compliant with the ARRIVE 2.0 guidelines for animal research reporting, were associated with human clinical results. ARRIVE 2.0 is a set of guidelines to improve the transparency and reproducibility of animal research. It is worth noting that early experimental studies on the effectiveness and safety of Dengvaxia reported breakthrough viremia in macaque models. The protection provided by the monoclonal antibody C10 was shown to mitigate antibody-dependent enhancement (ADE) in murine models, though it is not associated with the Dengvaxia vaccine. The further development of these animal models will be necessary to expedite the safe development of next-generation dengue vaccines (TAK-003, TV003/TV005), antivirals (favipiravir, sofosbuvir), and immune modulators (IL-6 and TNF-α blockers) for use in endemic areas.

登革热病毒（DENV）感染每年影响超过3.9亿人，其严重形式，包括登革出血热（DHF）和登革休克综合征（DSS），是抗体依赖性增强（ADE）、t细胞失调和细胞因子风暴的结果。本综述将人类免疫病理学结果与动物模型报告的结果相结合，以系统的方式巩固这些发现，以验证这些机制。AG129干扰素受体缺陷小鼠的研究一致表明，ade介导的病毒血症和血管渗漏。然而，人源化NSG-BLT小鼠在DENV异型攻击后再现了人类T细胞的典型抗原性。食蟹猴模型在生理学上与在接近人类循环条件下测试四价疫苗相关，并且使用饮食诱导的肥胖小鼠来表型代谢合并症对il -1β驱动的免疫病理的影响。使用标准化方法的数据，包括Evans蓝色外渗试验、多重细胞因子面板和符合动物研究报告ARRIVE 2.0指南的终点，与人类临床结果相关。ARRIVE 2.0是一套指导方针，旨在提高动物研究的透明度和可重复性。值得注意的是，早期关于登卡夏有效性和安全性的实验研究报道了在猕猴模型中突破病毒血症。在小鼠模型中，单克隆抗体C10提供的保护被证明可以减轻抗体依赖性增强（ADE），尽管它与登卡夏疫苗没有关联。有必要进一步开发这些动物模型，以加快安全开发用于流行地区的下一代登革热疫苗（TAK-003、TV003/TV005）、抗病毒药物（favipiravir、sofosbuvir）和免疫调节剂（IL-6和TNF-α阻断剂）。

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引用次数: 0

Development of a rapid and portable detection method for canine distemper virus based on CRISPR-Cas13a 基于CRISPR-Cas13a的犬瘟热病毒快速便携式检测方法的建立

IF 1.6 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS

Journal of virological methods

Pub Date : 2026-05-01 Epub Date: 2026-01-29 DOI: 10.1016/j.jviromet.2026.115355

Yuxuan Jiang , Zaixing Yang , Jiamei Yang , Yifan Li , Jiaqi Liu , Lili Zhao , Junwei Ge

Canine distemper virus (CDV) is a pathogenic microorganism that severely affects the respiratory, digestive, and nervous systems, causing multi-systemic symptoms. It infects nearly all terrestrial carnivores worldwide, particularly the Canidae and Mustelidae families, posing a serious threat to global socio-economic and public health security. Given the importance of etiological treatment and early diagnosis, developing novel detection methods with improved accuracy, rapidity, and user-friendliness is necessary for effective prevention and control of CDV infection. In this study, we established a novel testing method using recombinase-aid amplification (RAA) coupled with CRISPR-Cas13a and optimized the working concentration of CRISPR RNA (crRNA) and Cas13a for the lateral flow detection (LFD) of CDV. The RAA-CRISPR-Cas13a-LFD for CDV did not cross-react against other prevalent canine pathogens and the sensitivity can detect as little as 10² copies/μL of CDV cDNA plasmids. Additionally, combined with HUDSON this RAA-CRISPR-Cas13a-LFD method could be used to detect clinical samples within 1.5 h, with performance comparable to that of RT-PCR. The results for the RAA-CRISPR-Cas13a detection could be visualized using either fluorescence or lateral flow strips for in field-deployable viral diagnosis. Overall, our developed method showed good potential in point-of-care testing (POCT) to control and reduce the losses by CDV infection.

犬瘟热病毒（Canine犬瘟热病毒，CDV）是一种严重影响呼吸、消化和神经系统的病原微生物，可引起多系统症状。它感染世界上几乎所有的陆地食肉动物，特别是犬科和鼬科，对全球社会经济和公共卫生安全构成严重威胁。鉴于病原学治疗和早期诊断的重要性，开发新的检测方法提高准确性、快速性和用户友好性是有效预防和控制CDV感染的必要条件。本研究建立了重组酶辅助扩增（RAA）与CRISPR-Cas13a联用的检测方法，并优化了CRISPR RNA （crRNA）和Cas13a用于CDV侧流检测（LFD）的工作浓度。RAA-CRISPR-Cas13a-LFD检测CDV的灵敏度低至102拷贝/μL，与犬常见病原体无交叉反应。此外，结合HUDSON， RAA-CRISPR-Cas13a-LFD方法可在1.5 h内检测临床样品，性能与RT-PCR相当。RAA-CRISPR-Cas13a检测结果可以通过荧光或横向流动条进行可视化，用于现场部署病毒诊断。总的来说，我们开发的方法在即时检测（POCT）中显示出良好的潜力，可以控制和减少CDV感染造成的损失。

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引用次数: 0

Development of an ELISA using recombinant Dengue NS1 constructs to measure antibody titers and predict dengue disease outcome in a pediatric population in India 利用重组登革热NS1构建ELISA测定印度儿科人群的抗体滴度并预测登革热疾病结局。

IF 1.6 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS

Journal of virological methods

Pub Date : 2026-05-01 Epub Date: 2026-01-17 DOI: 10.1016/j.jviromet.2026.115351

Bharti Pathak , Aparna Chakravarty , N.Veena Rani , Anuja Krishnan

The majority of the antibody response in dengue infection targets the structural envelope (E) and membrane (M) proteins of the virus. However, vaccine development is challenging because antibodies against the E protein often show cross-reactivity, leading to antibody-dependent enhancement (ADE) and worsening of disease severity. Non-structural protein 1 (NS1), secreted during the acute phase of infection, has been implicated as a pathogenic factor that contributes to vascular permeability and haemorrhage in severe dengue. In this study, we aimed to investigate the role of NS1 antibodies in disease manifestation. Full-length NS1, along with its N-terminal (NS1A) and C-terminal (NS1B) regions, was expressed and purified to delineate the antibody response against different domains. Serum samples from 102 dengue-positive paediatric patients were analysed for antibody titres against these NS1 constructs using ELISA. Overall higher antibody titres against all three NS1 recombinant proteins were observed in mild cases compared to severe ones with most significant towards NS1A.These findings suggest that antibodies directed against NS1, particularly its N-terminal region, may have a protective role and could serve as a potential correlate of protection in dengue infection.

在登革热感染中，大多数抗体反应针对病毒的结构包膜(E)和膜(M)蛋白。然而，疫苗开发具有挑战性，因为针对E蛋白的抗体经常表现出交叉反应性，导致抗体依赖性增强（ADE）和疾病严重程度恶化。在感染急性期分泌的非结构蛋白1 （NS1）被认为是导致严重登革热患者血管通透性和出血的致病因素。在本研究中，我们旨在探讨NS1抗体在疾病表现中的作用。全长NS1及其n端（NS1A）和c端（NS1B）区域被表达和纯化，以描绘针对不同结构域的抗体反应。使用ELISA对102例登革热阳性患儿的血清样本进行了针对这些NS1构建体的抗体效价分析。与严重病例相比，在轻度病例中观察到针对所有三种NS1重组蛋白的抗体滴度总体较高，其中针对NS1A的抗体滴度最为显著。这些发现表明，针对NS1的抗体，特别是其n端区域，可能具有保护作用，并可能作为登革热感染保护的潜在关联。

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引用次数: 0

Development of a rapid detection kit for chicken infectious anemia virus nucleic acid (RPA-LFD method) 鸡传染性贫血病毒核酸（RPA-LFD）快速检测试剂盒的研制

IF 1.6 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS

Journal of virological methods

Pub Date : 2026-05-01 Epub Date: 2026-02-05 DOI: 10.1016/j.jviromet.2026.115363

Mei Tang , Lun Shu , Yifei Xiong , Qinmin Zhu , Yiwei Liu , Minfan Huang , Qiaoyang Teng , Chunxiu Yuan , Qinfang Liu , Xue Pan , Zhifei Zhang , Bangfeng Xu , Xiaona Shi , Minghao Yan , Dawei Yan , Zejun Li

Chicken infectious anemia virus (CIAV) is a major immunosuppressive pathogen that causes huge economic losses in the poultry industry. The virus persists in the host for an extended period, making it difficult to eliminate and highlighting the need for rapid and accurate detection methods. In this study, we developed a recombinase polymerase amplification-lateral flow dipstick (RPA-LFD) kit for on-site detection of CIAV. Optimal primers and probes were designed and screened based on conserved regions of the full-gene sequences of 251 CIAV strains, ensuring high specificity, with no cross-reactions with 12 other common avian viruses. It demonstrated high sensitivity with a minimum detection limit of 10 copies of CIAV DNA, achieving efficient detection in just 15 min. The kit also showed good repeatability and stability, with 100 % concordance with qPCR in detecting 50 field samples. The developed CIAV RPA-LFD kit provides a practical tool for the rapid detection of CIAV, suitable for both on-site and laboratory use.

鸡传染性贫血病毒（CIAV）是一种主要的免疫抑制病原体，给家禽业造成了巨大的经济损失。该病毒在宿主体内持续存在较长时间，使其难以消除，并突出表明需要快速和准确的检测方法。在本研究中，我们开发了一种用于CIAV现场检测的重组酶聚合酶扩增-横向流动试纸（RPA-LFD）试剂盒。根据251株CIAV病毒全基因序列的保守区设计和筛选最优引物和探针，具有较高的特异性，与其他12种常见禽流感病毒无交叉反应。该方法具有很高的灵敏度，最低检测限为10份CIAV DNA，仅在15分钟内即可实现高效检测。该试剂盒具有良好的重复性和稳定性，与qPCR检测50份野外样品的一致性为100%。开发的CIAV RPA-LFD试剂盒为CIAV的快速检测提供了实用的工具，适用于现场和实验室使用。

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引用次数: 0

Role of human cytomegalovirus in the pathogenesis of periapical lesions – A systematic review and meta-analysis 人巨细胞病毒在根尖周围病变发病机制中的作用——系统综述和荟萃分析。

IF 1.6 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS

Journal of virological methods

Pub Date : 2026-05-01 Epub Date: 2026-01-30 DOI: 10.1016/j.jviromet.2026.115359

Puja Chatterjee , Mala Kamboj , Anjali Narwal , Anju Devi , Adarsh Kumar

Background

Viruses, especially those of the Herpesviridae family, are increasingly linked to the pathogenesis of periapical lesions. Human cytomegalovirus (CMV), capable of lifelong latency and reactivation under stress or infection, can enhance proinflammatory cytokine release, stimulate osteoclast activity, and drive tissue destruction.

Objective

To evaluate the prevalence of CMV DNA and/or RNA in periapical lesions and to evaluate its potential role in their pathogenesis.

Materials and methods

A comprehensive search was performed using MEDLINE via PubMed, Google Scholar, Scopus, and ResearchGate up to April 2025, following PRISMA guidelines. Original studies that explored the role of CMV in periapical lesions were included. The risk of bias was evaluated with the modified Newcastle-Ottawa Scale, with results generated in Review Manager 5.4. Meta-analyses were carried out using OpenMeta Analyst.

Results

Thirty-one studies were included for qualitative analysis, of which 17 met eligibility for meta-analysis. The pooled prevalence of CMV in periapical lesions was 28 % (95 % CI: 17 %–40 %). Symptomatic lesions showed significantly higher CMV detection compared to asymptomatic cases (OR = 2.67, 95 % CI: 1.44–4.95). Subgroup analysis showed that CMV prevalence varied by detection method, with higher estimates using RT-PCR, indicating assay sensitivity as a major source of heterogeneity.

Conclusion

This review highlights that CMV was significantly associated with symptomatic and larger periapical lesions, likely contributing to disease progression via immune modulation, cytokine induction, and viral-bacterial interactions. Further research is warranted to clarify its pathogenic role and explore antiviral strategies in endodontic management.

背景：病毒，特别是疱疹病毒科的病毒，越来越多地与根尖周围病变的发病机制联系在一起。人巨细胞病毒（CMV）能够在应激或感染下终身潜伏和再激活，可以增强促炎细胞因子的释放，刺激破骨细胞活性，并驱动组织破坏。目的：评估CMV DNA和/或RNA在根尖周围病变中的流行程度，并评估其在根尖周围病变发病机制中的潜在作用。材料和方法：根据PRISMA指南，使用MEDLINE通过PubMed，谷歌Scholar， Scopus和ResearchGate进行了全面的检索，截止到2025年4月。探讨巨细胞病毒在根尖周围病变中的作用的原始研究包括在内。偏倚风险采用改良的Newcastle-Ottawa量表进行评估，结果在Review Manager 5.4中生成。meta分析使用OpenMeta Analyst进行。结果：31项研究纳入定性分析，其中17项符合荟萃分析的资格。根尖周围病变中巨细胞病毒的总患病率为28% （95% CI: 17%-40%）。有症状病变的CMV检出率明显高于无症状病变（OR = 2.67, 95% CI: 1.44-4.95）。亚组分析显示，CMV患病率因检测方法而异，RT-PCR的估计值较高，表明检测灵敏度是异质性的主要来源。结论：本综述强调巨细胞病毒与症状性和较大的根尖周围病变显著相关，可能通过免疫调节、细胞因子诱导和病毒-细菌相互作用促进疾病进展。需要进一步的研究来阐明其致病作用，并探索牙髓治疗中的抗病毒策略。

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引用次数: 0

Optimization of a GFP–PAC co-expression cassette–based purification system for efficient construction and rapid screening of recombinant HSV-1 基于GFP-PAC共表达盒的高效构建和快速筛选重组HSV-1纯化系统的优化

IF 1.6 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS

Journal of virological methods

Pub Date : 2026-05-01 Epub Date: 2026-02-11 DOI: 10.1016/j.jviromet.2026.115364

Mishar Kelishadi , Alijan Tabarraei , Kayhan Azadmanesh

Oncolytic viruses (OVs) constitute a promising class of cancer therapeutics engineered to selectively replicate in and lyse malignant cells while sparing normal tissues. Among these, recombinant Herpes Simplex Virus type 1 (HSV-1), generated through targeted deletion of immediate-early (IE) genes, has emerged as a leading candidate, with several vectors currently advancing to phase III clinical trials. Genetic alterations are typically introduced into the HSV-1 genome via homologous recombination using shuttle vectors; however, isolation of recombinant viruses from parental populations by plaque purification remains labor-intensive and technically demanding. This limitation primarily results from the low efficiency of homologous recombination and the frequent persistence of parental virus, which often exhibits a replicative advantage over recombinant variants. In this study, a novel purification protocol incorporating a GFP–PAC co-expression cassette was developed and optimized to enable simultaneous puromycin selection and fluorescence-based identification of recombinant viral constructs. This dual-selection approach significantly accelerates and simplifies the isolation of recombinant HSV-1, enhancing both efficiency and purity. The optimized protocol provides a robust, scalable strategy for generating recombinant HSV-1 with broad applicability in vaccine development, gene therapy, and fundamental virology research.

溶瘤病毒（OVs）是一类很有前途的癌症治疗药物，它被设计成选择性地在恶性细胞中复制和溶解，同时保留正常组织。其中，重组单纯疱疹病毒1型（HSV-1），通过靶向删除即时早期（IE）基因产生，已经成为主要的候选病毒，目前有几种载体正在进行III期临床试验。遗传改变通常通过使用穿梭载体的同源重组引入HSV-1基因组；然而，通过空斑纯化从亲本群体中分离重组病毒仍然是劳动密集型和技术要求高的。这种限制主要是由于同源重组的低效率和亲本病毒的频繁持续，亲本病毒往往表现出比重组变体更大的复制优势。在本研究中，开发并优化了一种包含GFP-PAC共表达盒的新型纯化方案，以同时进行嘌呤霉素选择和基于荧光的重组病毒构建体鉴定。这种双重选择方法显著加快和简化了重组HSV-1的分离，提高了效率和纯度。优化后的方案为产生重组HSV-1提供了一个强大的、可扩展的策略，在疫苗开发、基因治疗和基础病毒学研究中具有广泛的适用性。

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引用次数: 0