CRISPR/Cas12 System-Based Assay for Rapid, Sensitive Detection of Rotavirus in Food Samples.

IF 1.9 2区 农林科学 Q3 FOOD SCIENCE & TECHNOLOGY
Shirui Gou, Yan Liu, Qianqian Li, Jielin Yang, Long Qiu, Yu Zhao
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Abstract

Foodborne viruses have become an important threat to food safety and human health. Among the foodborne viruses, group A rotavirus is the most important pathogen of diarrhea in autumn and winter. The field detection of rotavirus is crucial for the early control of infection and patient management. Quantitative real-time reverse transcription-polymerase chain reaction is the most widely used in virus detection. However, the technique relies on high-cost instruments and trained personnel, which limit its use in field detection. In this study, we developed accurate, realizable, and simple detection methods by combining optimized CRISPR (clustered regularly interspaced short palindromic repeats) Cas12 and reverse transcription loop-mediated isothermal amplification (RT-LAMP) (reverse transcription loop-mediated isothermal amplification) to reduce the requirements for temperature control and costly real-time fluorescence polymerase chain reaction instruments. We investigated two nucleic acid detection systems combining RT-LAMP with CRISPR Cas12a and RT-LAMP with CRISPR Cas12b and compared them with reverse transcription-quantitative polymerase chain reaction. The resulting detection system only needs a reaction temperature and in single tube to react for 60 min with the detection sensitivity of 38 copies/μL. Overall, this study developed an innovative method for the rapid detection of rotavirus in food samples, which will help to effectively identify food contaminated by pathogens and prevent human infections and economic losses caused by disease outbreaks.

基于 CRISPR/Cas12 系统的检测方法,用于快速、灵敏地检测食品样本中的轮状病毒。
食源性病毒已成为食品安全和人类健康的重要威胁。在食源性病毒中,A 组轮状病毒是秋冬季腹泻的最主要病原体。轮状病毒的现场检测对早期控制感染和患者管理至关重要。定量实时反转录聚合酶链反应是目前应用最广泛的病毒检测方法。然而,该技术依赖于高成本的仪器和训练有素的人员,这限制了其在现场检测中的应用。在本研究中,我们将优化的 CRISPR(簇状规则间距短回文重复序列)Cas12 和反转录环介导等温扩增(RT-LAMP)(reverse transcription loop-mediated isothermal amplification)结合起来,开发出了准确、可实现且简单的检测方法,从而降低了对温度控制和昂贵的实时荧光聚合酶链反应仪器的要求。我们研究了结合 RT-LAMP 与 CRISPR Cas12a 和 RT-LAMP 与 CRISPR Cas12b 的两种核酸检测系统,并与反转录定量聚合酶链反应进行了比较。结果表明,该检测系统只需一个反应温度,在单管中反应 60 分钟,检测灵敏度为 38 个拷贝/μL。总之,本研究开发了一种创新的快速检测食品样品中轮状病毒的方法,有助于有效识别被病原体污染的食品,防止疾病爆发造成的人员感染和经济损失。
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来源期刊
Foodborne pathogens and disease
Foodborne pathogens and disease 医学-食品科技
CiteScore
5.30
自引率
3.60%
发文量
80
审稿时长
1 months
期刊介绍: Foodborne Pathogens and Disease is one of the most inclusive scientific publications on the many disciplines that contribute to food safety. Spanning an array of issues from "farm-to-fork," the Journal bridges the gap between science and policy to reduce the burden of foodborne illness worldwide. Foodborne Pathogens and Disease coverage includes: Agroterrorism Safety of organically grown and genetically modified foods Emerging pathogens Emergence of drug resistance Methods and technology for rapid and accurate detection Strategies to destroy or control foodborne pathogens Novel strategies for the prevention and control of plant and animal diseases that impact food safety Biosecurity issues and the implications of new regulatory guidelines Impact of changing lifestyles and consumer demands on food safety.
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