Development and evaluation of three multienzyme isothermal rapid amplification assays for fowl adenovirus serotype 4.

IF 3.8 1区 农林科学 Q1 AGRICULTURE, DAIRY & ANIMAL SCIENCE
Yin Dai, Yueyi Zhong, Fazhi Xu, Siqin Gu, Huiqin Zhou, Jieru Wang, Dongdong Yin, Lei Yin, Xuehuai Shen, Xiaocheng Pan, Xuelan Liu
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引用次数: 0

Abstract

Fowl adenovirus serotype 4 (FAdV-4) is the main causative agent of hydropericardium hepatitis syndrome (HHS), which has resulted in huge economic losses to the poultry industry in recent years. Hence, a rapid and simple visual detection method is needed for identification of FAdV-4. In this study, three multienzyme isothermal rapid amplification (MIRA) assays, basic MIRA, MIRA-qPCR and MIRA-LFD were developed for detection of FAdV-4. The amplification primers and reaction conditions were optimized, and the specificity and sensitivity of the assays were evaluated. The MIRA assays were specific for FAdV-4 with no cross-reaction with novel goose astrovirus, H9 subtype avian influenza virus, duck enteritis virus, Muscovy duck reovirus, or duck circovirus. The basic MIRA assay required only one primer pair and the reaction can be completed within 30 min at 36 °C. The MIRA-qPCR and MIRA-LFD assays were completed in 20 min with a minimum detection limit of 1 × 101 copies/μL and 1 × 102 copies/μL, respectively. The results of the MIRA-LFD assay can be observed directly with the naked eye, omitting the need for specialized instruments. The positive rate of three proposed MIRA assays were consistent with that of the conventional PCR assay. The MIRA assays are simple, rapid, and effective diagnostic tools for field detection of FAdV-4.

针对鸡腺病毒血清 4 型的三种多酶等温快速扩增检测方法的开发与评估。
鸡腺病毒血清型 4(FAdV-4)是心包积水肝炎综合征(HHS)的主要病原体,近年来给家禽业造成了巨大的经济损失。因此,需要一种快速、简单的肉眼检测方法来鉴定 FAdV-4。本研究开发了三种多酶等温快速扩增(MIRA)检测方法,即基本 MIRA、MIRA-qPCR 和 MIRA-LFD,用于检测 FAdV-4。对扩增引物和反应条件进行了优化,并对检测的特异性和灵敏度进行了评估。MIRA 检测法对 FAdV-4 具有特异性,与新型鹅天鹅病毒、H9 亚型禽流感病毒、鸭肠炎病毒、穆斯科维鸭再病毒或鸭圆环病毒没有交叉反应。基本的 MIRA 检测只需一对引物,反应可在 36 °C 下 30 分钟内完成。MIRA-qPCR 和 MIRA-LFD 检测在 20 分钟内完成,最低检测限分别为 1 × 101 拷贝/μL 和 1 × 102 拷贝/μL。MIRA-LFD 检测结果可直接用肉眼观察,无需专业仪器。三种拟议的 MIRA 检测法的阳性率与传统 PCR 检测法一致。MIRA 检测法是现场检测 FAdV-4 的简单、快速和有效的诊断工具。
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来源期刊
Poultry Science
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
0
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.
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