A novel biosensor for detecting V. parahaemolyticus based on cascade signal amplification of CRISPR/Cas14a and Exo III

IF 7 1区农林科学 Q1 FOOD SCIENCE & TECHNOLOGY

Food Control Pub Date : 2025-01-01 Epub Date: 2024-08-15 DOI:10.1016/j.foodcont.2024.110788

Jiahong Wang , He Sun , Yuhan Gao , Shengjun Bu , Zebin Zhang , Chen Wang , Hongyi Zhang , Wenhui Zhang , Jiayu Wan

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引用次数: 0

Abstract

Vibrio parahaemolyticus（V. parahaemolyticus）is a widespread food-borne pathogen in seafood, which poses a serious threat to human health and public safety. However, sensor platform projects that do not require complex sample processing are rare. Hence, we based on CRISPR/Cas14a and Exonuclease III (Exo III) to build a novel fluorescent biosensor that did not require nucleic acid extraction and amplification. A locked single-stranded DNA probe to a target-pathogen specific aptamer was designed to be opened as free single-stranded DNA (ssDNA) upon aptamer binding to the target pathogen. Exo III then cleaves hairpin B (HP B) from the 3′-terminus, releasing ssDNA probe A and ssDNA C. Then, ssDNA probe A hybridizes with another HP B to start a new cycle, and ssDNA C acts as a substrate to activate Cas14a/sgRNA trans-cleavage activity. By exploiting this novel design principle, we detected V. parahaemolyticus over a wide linear range with a limit of detection (LOD) of 6 CFU/mL.

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基于 CRISPR/Cas14a 和 Exo III 级联信号放大技术的新型副溶血性弧菌检测生物传感器

副溶血性弧菌（V. parahaemolyticus）是一种广泛存在于海产品中的食源性病原体，对人类健康和公共安全构成严重威胁。然而，不需要复杂样本处理的传感器平台项目并不多见。因此，我们基于CRISPR/Cas14a和Exonuclease III（Exo III）构建了一种无需核酸提取和扩增的新型荧光生物传感器。与目标病原体特异性适配体相连的锁定单链 DNA 探针被设计成在适配体与目标病原体结合后打开的自由单链 DNA（ssDNA）。然后，ssDNA探针A与另一个HP B杂交，开始一个新的循环，ssDNA C作为底物激活Cas14a/sgRNA反式裂解活性。利用这种新颖的设计原理，我们在很宽的线性范围内检测到了副溶血病毒，检测限（LOD）为 6 CFU/mL。

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

Food Control 工程技术-食品科技

CiteScore

12.20

自引率

6.70%

发文量

758

审稿时长

33 days

期刊介绍： Food Control is an international journal that provides essential information for those involved in food safety and process control. Food Control covers the below areas that relate to food process control or to food safety of human foods: • Microbial food safety and antimicrobial systems • Mycotoxins • Hazard analysis, HACCP and food safety objectives • Risk assessment, including microbial and chemical hazards • Quality assurance • Good manufacturing practices • Food process systems design and control • Food Packaging technology and materials in contact with foods • Rapid methods of analysis and detection, including sensor technology • Codes of practice, legislation and international harmonization • Consumer issues • Education, training and research needs. The scope of Food Control is comprehensive and includes original research papers, authoritative reviews, short communications, comment articles that report on new developments in food control, and position papers.