研究说明：通过基于 TaqMan 探针的多重 qPCR 同时检测 GPV、H5 AIV 和 GoAstV。

IF 3.8 1区农林科学 Q1 AGRICULTURE, DAIRY & ANIMAL SCIENCE

Poultry Science Pub Date : 2024-11-05 DOI:10.1016/j.psj.2024.104511

Xiaoquan Wang , Miao Cai , Xiaolong Lu , Qianqian Xu , Yanhong Wang , Wenhao Yang , Kaituo Liu , Ruyi Gao , Yu Chen , Jiao Hu , Min Gu , Shunlin Hu , Xiufan Liu , Xiaowen Liu

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引用次数: 0

摘要

鹅细小病毒（GPV）、H5 亚型禽流感病毒（AIV）和鹅星状病毒（GoAstV）感染的地方性流行状况继续对中国的家禽业造成严重破坏。尽管如此，分子诊断技术的应用仍存在明显差距。这项调查描述了一种多重定性聚合酶链反应（qPCR）检测方法的开发过程，该方法能够同时检测 GPV、H5 AIV 和 GoAstV，而且与其他禽类病毒病原体没有交叉反应。该检测方法对 GPV 和 GoAstV 的检测阈值为 10 拷贝/μL，对 H5 AIV 的检测阈值为 1 拷贝/μL。测定内和测定间的变异系数均小于 3.0%，表明测定批次内和测定批次间的重复性很高。利用这种多重 qPCR 检测方法，对一批 60 份临床样本进行了分析，以评估其实用性。检测到的 GoAstV、GPV 和 H5 AIV 感染率分别为 35.0%（21/60）、21.7%（13/60）和 15.0%（9/60）。还发现了并发感染，GPV + GoAstV、GPV + H5 AIV、GoAstV + H5 AIV 和 GPV + GoAstV + H5 AIV 的感染率分别为 6.7%（4/60）、3.3%（2/60）、3.3%（2/60）和 3.3%（2/60）。所开发的多重 qPCR 检测法的诊断吻合率与传统 PCR 技术相当。这种新型检测方法是一种快速、高效、特异和灵敏的工具，可用于检测流行的鹅病毒，从而加强疾病管理策略和流行病学监测工作。

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Research note: Simultaneous detection of GPV, H5 AIV, and GoAstV via TaqMan probe-based multiplex qPCR

The endemic status of goose parvovirus (GPV), H5 subtype avian influenza virus (AIV), and goose astrovirus (GoAstV) infections continues to devastate the poultry industry in China. Despite this, there exists a notable gap in the application of molecular diagnostic techniques. This investigation described the development of a multiplex qualitative polymerase chain reaction (qPCR) assay capable of concurrently detecting GPV, H5 AIV, and GoAstV, with no cross-reactivity observed with other avian viral pathogens. The assay exhibited a detection threshold of 10 copies/μL for both GPV and GoAstV, and 1 copy/μL for H5 AIV. The intra- and inter-assay coefficients of variation were < 3.0%, signifying high repeatability within and across assay batches. Utilizing this multiplex qPCR assay, a batch of 60 clinical samples was analyzed to assess its practical utility. The detected prevalence rates for GoAstV, GPV, and H5 AIV were 35.0% (21/60), 21.7% (13/60), and 15.0% (9/60), respectively. Concurrent infections were also identified, with rates for GPV + GoAstV, GPV + H5 AIV, GoAstV + H5 AIV, and GPV + GoAstV + H5 AIV being 6.7% (4/60), 3.3% (2/60), 3.3% (2/60), and 3.3% (2/60), respectively. The developed multiplex qPCR assay exhibited a diagnostic concordance rate equivalent to that of traditional PCR techniques. This novel assay serves as a rapid, efficient, specific, and sensitive tool for the detection of prevalent goose viruses, thereby enhancing disease management strategies and epidemiological monitoring efforts.

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来源期刊

Poultry Science 农林科学-奶制品与动物科学

CiteScore

7.60

自引率

15.90%

发文量

审稿时长

94 days

期刊介绍： First self-published in 1921, Poultry Science is an internationally renowned monthly journal, known as the authoritative source for a broad range of poultry information and high-caliber research. The journal plays a pivotal role in the dissemination of preeminent poultry-related knowledge across all disciplines. As of January 2020, Poultry Science will become an Open Access journal with no subscription charges, meaning authors who publish here can make their research immediately, permanently, and freely accessible worldwide while retaining copyright to their work. Papers submitted for publication after October 1, 2019 will be published as Open Access papers. An international journal, Poultry Science publishes original papers, research notes, symposium papers, and reviews of basic science as applied to poultry. This authoritative source of poultry information is consistently ranked by ISI Impact Factor as one of the top 10 agriculture, dairy and animal science journals to deliver high-caliber research. Currently it is the highest-ranked (by Impact Factor and Eigenfactor) journal dedicated to publishing poultry research. Subject areas include breeding, genetics, education, production, management, environment, health, behavior, welfare, immunology, molecular biology, metabolism, nutrition, physiology, reproduction, processing, and products.