与RPA-CRISPR/Cas12a合的微流体生物传感器,可在现场进行对Vibrio parahaemolyticus的敏感量化
Jiehong Fang1, Qi Chen1, Meijuan Ran1
1Key Laboratory of Specialty Agri-products Quality and Hazard Controlling Technology of Zhejiang Province, College of Life Sciences, China Jiliang University, Hangzhou, Zhejiang, 310018, China.
Biosensors & bioelectronics
|December 23, 2025
概括
一种新方法可以使用同热放大和CRISPR技术对Vibrio parahaemolyticus进行可靠的现场定量. 这种低成本的测定方法通过提供准确的病原体检测,而无需复杂的设备来提高海鲜安全和水产养殖.
科学领域:
- 生物技术是生物技术.
- 食品安全 食品安全
- 水产养殖健康 水产养殖健康
背景情况:
- 紫外线病毒 (Vibrio parahaemolyticus) 是一个重要的病原体,导致海鲜传播的疾病和水生动物疾病.
- 准确的现场检测对于管理公共卫生风险和水产养殖至关重要.
- 由于复杂的仪器,现有的方法,如qPCR在现场应用中受到限制.
研究的目的:
- 开发一种强大的,可在现场部署的定量异热试验,用于检测Vibrio parahaemolyticus.
- 为了实现基于标准曲线的量化,在运行时进行校准和差异控制.
- 建立基于CRISPR的定量RPA测试的可转移方法.
主要方法:
- 一个离心式微流体系统,集成反应体积锁定和同时生成标准曲线.
- 一个单复合酶聚合酶放大 (RPA) -CRISPR/Cas12a试验,具有优化的crRNA和反应条件.
- 同时分析标准和样本以减少变异性.
主要成果:
- 达到6.08副本/μL的检测极限和10^0-10^4副本/μL的线性定量范围 (R2 > 0.96).
- 已证明的性能与qPCR (AUC = 0.984) 相比,具有可接受的试验内和试验间的可变性 (RSD 2.63-6.07%).
- 在加和真实海鲜样本上进行验证,显示可靠的现场定量,每次测试约为3.30美元.
结论:
- 拟议的方法框架允许在现场条件下使用同热放大方法进行可靠的定量检测.
- 集成的微流体平台和RPA-CRISPR/Cas12a试验为病原体检测提供了低成本,准确的解决方案.
- 这项工作促进了水产养殖和食品安全的现场部署诊断,解决了关键的公共卫生问题.
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