Journal of virological methods最新文献_第2页

Fully automated sample-to-answer PCR detection of severe fever with thrombocytopenia syndrome virus in cat serum and whole blood 猫血清和全血中发热伴血小板减少综合征病毒的全自动PCR检测。

IF 1.6 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS

Journal of virological methods

Pub Date : 2026-01-24 DOI: 10.1016/j.jviromet.2026.115353

Shun Sasaki , Keita Ishijima , Ken Maeda , Marifu Yamagishi-Kondoh

Severe fever with thrombocytopenia syndrome virus (SFTSV) is a tick-borne pathogen that causes fatal hemorrhagic disease in humans and animals. In addition to transmission via tick bites, infection can occur through contact with body fluids from infected individuals or animals. This study evaluated the POCKIT™ Central Nucleic Acid Analyzer (POCKIT Central), a fully automated system integrating nucleic acid extraction and insulated isothermal PCR as a rapid diagnostic tool to aid in preventing the spread of SFTSV infection in veterinary medicine. Analytical sensitivity was assessed using four viral strains; all were detectable, although some exhibited reduced sensitivity. In feline whole blood or serum spiked with SFTSV, the system demonstrated sufficient sensitivity for primary diagnosis of cats showing clinical signs of SFTS. Results of analyses of clinical serum samples using POCKIT Central perfectly matched those obtained using conventional reverse transcription–polymerase chain reaction. The POCKIT Central system can detect SFTSV directly in whole blood or serum within 85 min, without requiring complex manual procedures, thus enabling rapid diagnosis and timely therapeutic intervention in veterinary settings, and it also has the potential for application to other animal species.

发热伴血小板减少综合征病毒（SFTSV）是一种蜱传病原体，可在人和动物中引起致命的出血性疾病。除了通过蜱叮咬传播外，感染还可通过接触受感染个体或动物的体液发生。本研究评估POCKIT™中心核酸分析仪（POCKIT Central），这是一种全自动系统，集成了核酸提取和绝缘等温PCR，作为一种快速诊断工具，有助于预防SFTSV感染在兽药中的传播。使用四种病毒株评估分析敏感性；所有的都是可检测的，尽管有些表现出较低的灵敏度。在添加了SFTSV的猫全血或血清中，该系统对出现SFTS临床症状的猫的初步诊断显示出足够的敏感性。POCKIT Central对临床血清样本的分析结果与常规逆转录聚合酶链反应的结果完全吻合。POCKIT Central系统可以在85分钟内直接检测全血或血清中的SFTSV，无需复杂的人工操作，从而在兽医环境中实现快速诊断和及时的治疗干预，并且它也具有应用于其他动物物种的潜力。

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引用次数: 0

Comparative analysis of beet western yellows virus detection methods: Development and validation of a novel real-time RT-PCR assay also targeting beet leaf yellowing virus 甜菜西部黄病毒检测方法的比较分析：一种新的实时RT-PCR检测方法的建立和验证，也针对甜菜叶黄病毒。

IF 1.6 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS

Journal of virological methods

Pub Date : 2026-01-22 DOI: 10.1016/j.jviromet.2026.115352

Hellin Pierre , Everaert Ellen , Demonty Elisabeth , Richet Pauline , Muhovski Yordan , De Jonghe Kris , Hautier Louis , Steyer Stéphan

Beet western yellows virus (BWYV – Polerovirus BWYV) is a polerovirus associated with yellowing and stunting symptoms in sugar beet and several other crops. Its close relationship with other beet-infecting poleroviruses, has long complicated accurate diagnosis due to serological cross-reactivity and limited molecular data. To overcome these issues, we developed a novel real-time RT-PCR assay enabling the specific and sensitive detection of BWYV and beet leaf yellowing virus (BLYV – Polerovirus BLYV). The assay was designed from a curated genomic dataset to ensure inclusivity and tested in parallel with validated reference methods, including a semi-generic ELISA and a generic polerovirus RT-PCR. Comprehensive validation following the EPPO PM 7/98(5) standard confirmed the method’s high analytical sensitivity, specificity, repeatability, and reproducibility. The assay proved robust to methodological variations and compatible with multiplex diagnostic workflows. Compared with existing approaches, it offers faster, more reliable detection and eliminates cross-reactivity with non-target poleroviruses. This method provides a valuable tool for accurate BWYV surveillance and supports improved management of beet virus yellows in agricultural and phytosanitary contexts.

甜菜西部黄病毒（BWYV - Polerovirus BWYV）是一种与甜菜和其他几种作物的发黄和发育迟缓症状相关的Polerovirus。它与其他感染甜菜的多极病毒关系密切，由于血清学交叉反应性和有限的分子数据，长期以来使准确诊断复杂化。为了克服这些问题，我们开发了一种新的实时RT-PCR检测BWYV和甜菜叶变黄病毒（BLYV - Polerovirus BLYV）的特异性和敏感性。该检测是根据精心设计的基因组数据集设计的，以确保包容性，并与经过验证的参考方法（包括半通用ELISA和通用多病毒RT-PCR）并行测试。按照EPPO PM 7/98(5)标准进行全面验证，证实该方法具有较高的分析灵敏度、特异性、重复性和再现性。该试验证明了方法变化的稳健性，并与多种诊断工作流程兼容。与现有方法相比，它提供了更快、更可靠的检测，并消除了与非靶标多病毒的交叉反应。该方法为准确监测BWYV提供了有价值的工具，并支持在农业和植物检疫环境下改进甜菜病毒黄的管理。

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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 : 2026-01-17 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

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 : 2026-01-16 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

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 : 2026-01-12 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

Corrigendum to "Targeted enrichment of elephant endotheliotropic herpesvirus for complete genome sequencing in elephants" [J. Virol. Methods 338 (2025), 115230]. 大象嗜内皮疱疹病毒全基因组测序的靶向富集[J]。性研究。方法338(2025),115230]。

IF 1.6 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS

Journal of virological methods

Pub Date : 2026-01-09 DOI: 10.1016/j.jviromet.2026.115348

Kirtika Sharma, Kg Sai Balaji, Gaurav Kumar Sharma, Abhijit M Pawde, Sonalika Mahajan, Ravikant Agrawal, Pracheta Janmeda, Karikalan Mathesh

引用次数: 0

Genetic influence of IFN-γ gene polymorphisms on hepatitis C progression and recovery IFN-γ基因多态性对丙型肝炎进展和恢复的遗传影响

IF 1.6 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS

Journal of virological methods

Pub Date : 2026-01-06 DOI: 10.1016/j.jviromet.2026.115337

Khair Rafiq, Sanaullah Khan, Muhammad Bar Khan, Muhammad Yaqoob, Muhammad Adnan

Background

Hepatitis C virus (HCV) infection is a significant health concern that leads to liver complications. The interferon gamma (INF-γ) and interferon gamma receptor-1 (IFN-γR1) genes play role in viral infections. This study aims to analyse the genetic influences of IFN-γ and IFN-γR1 genes polymorphisms on HCV infection outcomes.

Methodology

This cross-sectional study comprised 320 subjects, were classified in to two groups; 160 HCV patients and 160 healthy controls. The HCV patients included 50 subjects with spontaneous viral clearance (SVC), 80 patients with chronic hepatitis C (CHC) and 30 subjects with sustained virological responder (SVR). RNA and DNA were extracted from blood samples. HCV detection and genotyping was performed through PCR. The intron-1 of INF-γ and promoter of IFN-γR1 genes were amplified through PCR and sequenced through Sanger’s method.

Results

The frequency of TT, AT and AA genotypes of IFN-γ at + 874 in HCV positive patients was 62(38.75 %), 64(40 %) and 34(21.5 %); in healthy individuals 73(46.0 %), 61(38.12 %) and 26(16.0 %) (p = 0.153). Genotype AT was significantly high in CHC patients 45(57.0 %) while the TT was considerably high in SVC 27(54.0 %) (p < 0.0001). The frequency of TT, TC, and CC genotypes of IFN-γR1 at −56 was 41(25.64 %), 56(35.0 %), and 63(39.36 %) in HCV patients, while 61(38.13 %), 33(20.63 %) and 66(41.25) in healthy controls (p = 0.0001). The TC genotype was detected high in CHC 34(42.60 %) while the CC was observed more in SVC 24(48.0 %) (P < 0.0001).

Conclusion

IFN-γ AT and IFN-γR1 TC genotypes are associated with Hepatitis C susceptibility, while IFN-γ TT and IFN-γR1 CC may influence recovery.

丙型肝炎病毒（HCV）感染是导致肝脏并发症的重要健康问题。干扰素γ (INF-γ)和干扰素γ受体-1 （IFN-γ r1）基因在病毒感染中起作用。本研究旨在分析IFN-γ和IFN-γ r1基因多态性对HCV感染结局的遗传影响。方法横断面研究共纳入320名受试者，分为两组；160名HCV患者和160名健康对照者。HCV患者包括50例自发性病毒清除（SVC）患者，80例慢性丙型肝炎（CHC）患者和30例持续病毒学应答（SVR）患者。从血样中提取RNA和DNA。采用PCR进行HCV检测和基因分型。通过PCR扩增IFN-γ内含子-1和IFN-γ r1基因启动子，并通过Sanger法测序。结果HCV阳性患者IFN-γ + 874的TT、AT和AA基因型频率分别为62（38.75 %）、64（40 %）和34(21.5 %)；73年健康个体(46.0 %),61(38.12 %)和26(16.0 %)(p = 0.153)。基因型AT在CHC患者45中显著高（57.0% %），而TT在SVC患者27中显著高（54.0% %）（p <； 0.0001）。HCV患者TT、TC和CC基因型分别为41（25.64 %）、56（35.0 %）和63(39.36 %)，健康对照组分别为61（38.13 %）、33（20.63 %）和66(41.25)（p = 0.0001）。TC基因型在CHC 34中较高（42.60 %），而CC基因型在SVC 24中较多（48.0 %）（P <； 0.0001）。结论IFN-γ AT和IFN-γ r1 TC基因型与丙型肝炎易感性相关，而IFN-γ TT和IFN-γ r1 CC可能影响康复。

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引用次数: 0

Development and application of a real-time RT-PCR assay for the specific detection of influenza D virus D型流感病毒特异性实时RT-PCR检测方法的建立与应用

IF 1.6 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS

Journal of virological methods

Pub Date : 2026-01-05 DOI: 10.1016/j.jviromet.2026.115338

Jinfeng Wang , Ruiwen Li , Libing Liu , Jieshi Yu , Ning Li , Xincheng Ji , Yue Yang , Xiaoxia Sun , Wanzhe Yuan , Jianchang Wang

Since the influenza D virus (IDV) was first isolated from a swine exhibiting respiratory disease symptoms in the United States in 2011, it has been detected in various animals across more than 20 countries worldwide, including cattle, pigs, goats, sheep, and camels. IDV has emerged as a significant pathogen associated with bovine respiratory disease complex (BRDC). In this study, specific primers and a TaqMan probe were designed based on the conserved region in the PB1 gene to develop a highly specific and efficient real-time RT-PCR assay (RT-qPCR) for IDV. The RT-qPCR assay demonstrated excellent specificity for IDV, without cross-reactions with other common BRDC-associated pathogens. Using in vitro transcribed PB1 RNA as a template, the assay's limit of detection was established at 100 copies/reaction. Furthermore, the developed RT-qPCR assay was evaluated using 417 cattle nasal swab samples collected from 10 different regions in Hebei Province, China, between 2021 and 2022. The results showed that 33 samples (7.91 %) tested positive for IDV. The IDV HEF gene was amplified and sequenced in these 33 positive samples, yielding 10 HEF gene sequences that all belonged to the D/Yama2019 lineage, with nucleotide homology ranging from 98.7 % to 99.1 %. This study successfully developed an RT-qPCR assay for the specific and sensitive detection of IDV, which performed well on the cattle nasal samples. Additionally, it is the first to demonstrate the presence of IDV in cattle herds in Hebei Province, China.

自2011年在美国首次从出现呼吸道疾病症状的猪身上分离出D型流感病毒（IDV）以来，该病毒已在全球20多个国家的各种动物身上被发现，包括牛、猪、山羊、绵羊和骆驼。IDV已成为与牛呼吸道疾病复合体（BRDC）相关的重要病原体。本研究基于PB1基因的保守区设计了特异性引物和TaqMan探针，建立了IDV高特异性、高效的实时RT-PCR检测方法。RT-qPCR检测显示IDV具有良好的特异性，与其他常见brdc相关病原体无交叉反应。以体外转录的PB1 RNA为模板，测定的检测限为100拷贝/反应。此外，利用2021年至2022年间从中国河北省10个不同地区收集的417份牛鼻拭子样本，对所开发的RT-qPCR方法进行了评估。结果显示，33份样本（7.91%）检测出IDV阳性。在33份阳性样本中扩增并测序IDV HEF基因，得到10个HEF基因序列，均属于D/Yama2019谱系，核苷酸同源性为98.7% ~ 99.1%。本研究成功建立了一种特异、灵敏的检测IDV的RT-qPCR方法，该方法在牛鼻样品中表现良好。此外，这是中国河北省首次证实牛群中存在IDV。

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引用次数: 0

Corrigendum to “An alternative model for HEV infection in the HepaRG cell line” [J. Virol. Methods 340 (2026) 115307] “HepaRG细胞系中HEV感染的另一种模型”的更正[J]。性研究。方法340(2026)115307。

IF 1.6 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS

Journal of virological methods

Pub Date : 2025-12-24 DOI: 10.1016/j.jviromet.2025.115334

Stacy Gellenoncourt , Marie Pellerin , Aïlona Marcadet-Hauss , Roxanne Fouillé , Michel Rivoire , Guillaume Passot , Julie Lucifora , David Durantel , Nicole Pavio , Virginie Doceul

引用次数: 0

Evaluation of dried blood spot testing for serological monitoring of epizootic and zoonotic pathogens in domestic pigs 家猪兽疫和人畜共患病原体血清学监测中干血斑点试验的评价。

IF 1.6 4区医学 Q3 BIOCHEMICAL RESEARCH METHODS

Journal of virological methods

Pub Date : 2025-12-24 DOI: 10.1016/j.jviromet.2025.115336

Ranja Steinhauer , Eric Kübler , Stefan Gaugler , Cornel Fraefel , Julia Lechmann

Dried blood spots (DBS) constitute a stable, cost-efficient sampling matrix that can be collected in a minimally invasive manner. Although widely adopted in human medicine, their use in veterinary diagnostics remains limited. This study aimed to establish and validate DBS elution protocols for use in commercial ELISAs to detect antibodies against Hepatitis E virus (HEV), African swine fever virus (ASFV) and Aujeszky’s disease virus (ADV) in domestic pigs. DBS were prepared from EDTA blood, dried serum spots (DSS) from serum. Additional DBS samples were prepared after spiking blood from healthy pigs with antibodies. Various parameters were evaluated to establish the final elution protocols, i.e. number of disks, type and volume of elution buffer, incubation time of the disks in elution buffer, and volume of eluate used for detection. Once the final protocols were in place, for each pathogen 52 DBS were tested in three independent runs. The diagnostic performance was evaluated by comparing the ELISA results of DBS eluates with the corresponding serum or plasma samples. For HEV, only one out of 52 DBS samples qualitatively did not match the plasma result in any of the three runs. For ASFV, all 52 DBS samples matched the qualitative results of the corresponding liquid samples. For ADV, two samples yielded false negative results in all three runs. The results suggest that DBS represent a practical and reliable alternative to liquid blood samples for antibody detection in pigs. Further validation with field samples and large-scale testing is needed.

干血斑（DBS）构成了一种稳定、成本效益高的采样基质，可以以微创的方式采集。虽然在人类医学中被广泛采用，但它们在兽医诊断中的应用仍然有限。本研究旨在建立和验证DBS洗脱方案，用于商用elisa检测家猪戊型肝炎病毒（HEV）、非洲猪瘟病毒（ASFV）和奥杰斯基病病毒（ADV）的抗体。EDTA血制备DBS，血清干斑（DSS）制备DBS。在健康猪的血液中加入抗体后，制备了额外的DBS样本。评估各种参数以确定最终洗脱方案，包括：圆盘数、洗脱缓冲液的类型和体积、圆盘在洗脱缓冲液中的孵育时间、用于检测的洗脱液体积。一旦最终方案到位，每种病原体52个DBS在三个独立的运行中进行测试。通过比较DBS洗脱液的ELISA结果与相应的血清或血浆样品来评估诊断性能。对于HEV， 52个DBS样本中只有一个在三次运行中与血浆结果定性不匹配。对于ASFV， 52份DBS样品均与相应液体样品的定性结果相匹配。对于ADV，两个样本在所有三次运行中都产生了假阴性结果。结果表明，DBS是一种实用可靠的猪抗体检测方法，可替代液体血液样品。需要用现场样品和大规模试验进一步验证。

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引用次数: 0