On-Site Rapid and Specific Detection of Novel Emerging Goose Astrovirus Using a Nanobody-Based Colloidal Gold Immunochromatographic Strip

IF 3.5 2区农林科学 Q2 INFECTIOUS DISEASES

Transboundary and Emerging Diseases Pub Date : 2025-07-01 DOI:10.1155/tbed/6439771

Dan Wang, Yanhong Ji, Bin Liu, Lihuan Yan, Yi Ai, Jing Wang, Tao Jiang, Mi Lin, Qiyun Zhu

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引用次数: 0

Abstract

In recent years, outbreaks of gouty goose disease in goslings caused by a novel goose astrovirus (nGAstV) have occurred across major poultry-producing regions in Eastern China, with a mortality rate of approximately 50%. To date, there is a lack of rapid detection kits for early-stage disease control to reduce economic losses. In this study, two nanobodies (Nb-58 and Nb-60) that reacted with the nGAstV P2 protein were screened and identified using phage display technology and immunological experiments to develop colloidal gold immunochromatographic strips (CGISs) for nGAstV detection. CGIS did not cross-react with goose parvovirus (GPV), goose circovirus (GCV), H9N2 avian influenza virus (H9N2 AIV), Newcastle disease virus (NDV), avian leukemia virus (ALV), or infectious bursal disease virus (IBDV), indicating its strong specificity. The detection threshold of the CGIS was more than 640-fold diluted nGAstV (~10^1.57 TCID₅₀), which was comparable to a Ct value of 26.82, thereby showing high sensitivity. CGIS, with high stability, could be stored for at least 6 months at 25 and 4°C. The repeatability test showed that nGAstV could be readily detected in different CGIS batches. The coincidence rates of CGIS and conventional reverse transcription-polymerase chain reaction (RT-PCR) in 51 clinical tissue samples and 60 cloacal swab samples were 100% and 95.24%, respectively. In conclusion, the present study identified two specific Nbs and developed a reliable CGIS for the rapid detection of nGAstV in the field setting.

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基于纳米体胶体金免疫层析条带的新型鹅星状病毒现场快速特异性检测

近年来，由一种新型鹅星状病毒（nGAstV）引起的雏鹅痛风病在中国东部主要家禽产区暴发，死亡率约为50%。迄今为止，缺乏用于早期疾病控制以减少经济损失的快速检测试剂盒。本研究通过噬菌体展示技术和免疫学实验，筛选了与nGAstV P2蛋白发生反应的两种纳米小体Nb-58和Nb-60，并对其进行了鉴定，制备了用于nGAstV检测的胶体金免疫层析条（CGISs）。CGIS与鹅细小病毒（GPV）、鹅圆环病毒（GCV）、H9N2禽流感病毒（H9N2 AIV）、新城疫病毒（NDV）、禽白血病病毒（ALV）、传染性法氏囊病病毒（IBDV）均无交叉反应，特异性强。CGIS的检测阈值为稀释后的nGAstV的640倍以上（~101.57 TCID50），与26.82的Ct值相当，具有较高的灵敏度。CGIS稳定性高，可在25℃和4℃条件下保存至少6个月。重复性试验表明，nGAstV可在不同批次的CGIS中检测到。51份临床组织样本和60份肛肠拭子样本的CGIS与常规逆转录聚合酶链反应（RT-PCR）符合率分别为100%和95.24%。总之，本研究确定了两个特定的Nbs，并开发了一个可靠的CGIS，用于在现场环境中快速检测nGAstV。

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来源期刊

Transboundary and Emerging Diseases 农林科学-传染病学

CiteScore

8.90

自引率

9.30%

发文量

350

审稿时长

1 months

期刊介绍： Transboundary and Emerging Diseases brings together in one place the latest research on infectious diseases considered to hold the greatest economic threat to animals and humans worldwide. The journal provides a venue for global research on their diagnosis, prevention and management, and for papers on public health, pathogenesis, epidemiology, statistical modeling, diagnostics, biosecurity issues, genomics, vaccine development and rapid communication of new outbreaks. Papers should include timely research approaches using state-of-the-art technologies. The editors encourage papers adopting a science-based approach on socio-economic and environmental factors influencing the management of the bio-security threat posed by these diseases, including risk analysis and disease spread modeling. Preference will be given to communications focusing on novel science-based approaches to controlling transboundary and emerging diseases. The following topics are generally considered out-of-scope, but decisions are made on a case-by-case basis (for example, studies on cryptic wildlife populations, and those on potential species extinctions): Pathogen discovery: a common pathogen newly recognised in a specific country, or a new pathogen or genetic sequence for which there is little context about — or insights regarding — its emergence or spread. Prevalence estimation surveys and risk factor studies based on survey (rather than longitudinal) methodology, except when such studies are unique. Surveys of knowledge, attitudes and practices are within scope. Diagnostic test development if not accompanied by robust sensitivity and specificity estimation from field studies. Studies focused only on laboratory methods in which relevance to disease emergence and spread is not obvious or can not be inferred (“pure research” type studies). Narrative literature reviews which do not generate new knowledge. Systematic and scoping reviews, and meta-analyses are within scope.