Visual detection of fowl adenovirus serotype 4 via a portable CRISPR/Cas13a-based lateral flow assay.

IF 2.5 2区农林科学 Q1 VETERINARY SCIENCES Avian Pathology Pub Date : 2023-12-01 Epub Date: 2023-10-11 DOI:10.1080/03079457.2023.2254253

Dongdong Yin, Lei Yin, Jieru Wang, Yin Dai, Xuehuai Shen, Ruihong Zhao, Kezong Qi, Xiaocheng Pan

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Abstract

The widespread occurrence of fowl adenovirus serotype 4 (FAdV-4)-induced hepatitis-hydropericardium syndrome (HHS) has led to significant economic losses for the poultry industry. A sensitive, accurate, and practical FAdV-4 diagnostic approach is urgently required to limit the incidence of the disease. In the present study, a practical method for detecting FAdV-4 was developed using the CRISPR/Cas13a system and recombinase-aided amplification. The approach was based on 37°C isothermal detection with visible results being achieved. The detection limit of the target gene with this approach was only 10¹ copies/μl, making it very sensitive and specific. Clinical samples fared well when tested with the Cas13a detection method. For identifying FAdV-4, this novel detection approach was found to be sensitive, specific, and effective.RESEARCH HIGHLIGHTS First study using the CRISPR/Cas13a-based lateral flow detection assay for FAdV-4 detection.The results can be observed by the naked eye.The developed assay could provide an alternative tool for detection of FAdV-4 with minimal equipment.

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通过基于便携式CRISPR/Cas13a的侧流测定法目视检测家禽腺病毒血清型4。

家禽腺病毒血清型4（FAdV-4）诱导的肝炎-心包积液综合征（HHS）的广泛发生给家禽业带来了重大的经济损失。迫切需要一种灵敏、准确和实用的FAdV-4诊断方法来限制该疾病的发病率。在本研究中，使用CRISPR/Cas13a系统和重组酶辅助扩增开发了一种检测FAdV-4的实用方法。该方法基于37°C等温检测，取得了明显的结果。该方法对靶基因的检测限仅为101 拷贝数/μl，使其非常敏感和特异。当用Cas13a检测方法测试时，临床样本表现良好。对于识别FAdV-4，这种新的检测方法被发现是敏感、特异和有效的。研究亮点首次使用基于CRISPR/Cas13a的侧流检测法检测FAdV-4。结果可以用肉眼观察到。所开发的测定法可以用最少的设备提供检测FAdV-4的替代工具。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Avian Pathology 农林科学-兽医学

CiteScore

4.50

自引率

10.70%

发文量

审稿时长

1 months

期刊介绍： Avian Pathology is the official journal of the World Veterinary Poultry Association and, since its first publication in 1972, has been a leading international journal for poultry disease scientists. It publishes material relevant to the entire field of infectious and non-infectious diseases of poultry and other birds. Accepted manuscripts will contribute novel data of interest to an international readership and will add significantly to knowledge and understanding of diseases, old or new. Subject areas include pathology, diagnosis, detection and characterisation of pathogens, infections of possible zoonotic importance, epidemiology, innate and immune responses, vaccines, gene sequences, genetics in relation to disease and physiological and biochemical changes in response to disease. First and subsequent reports of well-recognized diseases within a country are not acceptable unless they also include substantial new information about the disease or pathogen. Manuscripts on wild or pet birds should describe disease or pathogens in a significant number of birds, recognizing/suggesting serious potential impact on that species or that the disease or pathogen is of demonstrable relevance to poultry. Manuscripts on food-borne microorganisms acquired during or after processing, and those that catalogue the occurrence or properties of microorganisms, are unlikely to be considered for publication in the absence of data linking them to avian disease.