Development of an RT-ERA-CRISPR/Cas12a assay for duck Tembusu virus

IF 4.2 1区农林科学 Q1 AGRICULTURE, DAIRY & ANIMAL SCIENCE

Poultry Science Pub Date : 2026-05-01 Epub Date: 2026-02-10 DOI:10.1016/j.psj.2026.106612

Zhanhong Huang , Yangbao Ding , Haocheng Xu , Qi Zhang , Xiyi Wang , Huamao Ding , Junsheng Zhang , Yufei Wang , Yingying Wang , Peirong Jiao

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引用次数: 0

Abstract

Duck Tembusu virus (DTMUV) has continued to threaten the duck industry in China since 2010. Therefore, the establishment of a rapid, specific, and sensitive method for the field detection of DTMUV is urgently needed. Herein, a reverse transcription enzymatic recombinase amplification (RT-ERA) assay was combined with the CRISPR/Cas12a system to target the DTMUV C gene. This assay exhibited high specificity, effectively distinguishing DTMUV from other common avian viruses. Its limit of detection reached 1 copy/μL DTMUV RNA. Moreover, this assay can be completed at 42 °C within 15 min using a thermostatic water bath. Additionally, we tested 30 clinical samples from infected ducks using this assay, and the results showed 100% concordance with SYBR green quantitative PCR results. In summary, this rapid, specific, and sensitive assay shows promising potential for DTMUV detection in the field.

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鸭坦布苏病毒RT-ERA-CRISPR/Cas12a检测方法的建立

自2010年以来，鸭坦布苏病毒（DTMUV）一直威胁着中国的养鸭业。因此，迫切需要建立一种快速、特异、灵敏的DTMUV现场检测方法。本文将逆转录酶重组酶扩增（RT-ERA）检测与CRISPR/Cas12a系统相结合，靶向DTMUV C基因。该方法具有高特异性，能有效地将DTMUV与其他常见禽流感病毒区分开来。其检测限达到1拷贝/μL DTMUV RNA。此外，该实验可以在42°C恒温水浴15分钟内完成。此外，我们用该方法检测了30份感染鸭的临床样本，结果与SYBR绿色定量PCR结果的一致性为100%。总之，这种快速、特异、灵敏的检测方法在DTMUV的现场检测中显示出很大的潜力。

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

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.