A CRISPR/Cas12a biosensor for portable and accessible detection of Salmonella typhimurium via multi-indicator pH millidisc colorimetry and smartphone imaging platform

IF 10.7 1区生物学 Q1 BIOPHYSICS

Biosensors and Bioelectronics Pub Date : 2025-05-20 DOI:10.1016/j.bios.2025.117611

Miaolin Duan , Guoqiang Li , Jingqi Shen , Ruitong Dai , Xingmin Li , Zunying Liu , Fei Jia

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引用次数: 0

Abstract

Conventional colorimetric CRISPR/Cas methods rely on a single chromogenic substrate and bulky and specialized signal detection instrument, which hinder their practical application. Herein, a portable and accessible CRISPR/Cas12a biosensor was for the first time reported to sensitively quantify Salmonella enterica serovar typhimurium (S. typhimurium), utilizing a multi-indicator pH millimeter disc (millidisc) for signal visualization, combined with a smartphone-based imaging platform for signal readout. The pH millidisc, composed of multiple indicators, possessed sensitive pH responsiveness and exhibited diverse color changes. The self-developed RGB mini-program, named DeepFood, was designed for portable smartphone use, featuring user-friendly operation and trend visualization for preliminary result analysis. Based on the RGB signal variation pattern with S. typhimurium, a distinct S_enh signal type was designed, enhancing the signal-to-noise ratio from 3.38 to 7.11. Compared to the R signal type, the S_enh signal type improved detection sensitivity by 36.23-fold (7.26 CFU/mL) in 0.01 M PBS buffer and 15.53-fold (1.41 × 10² CFU/mL) in chicken. The proposed biosensor offers significant improvements in detection sensitivity and practical applicability, with potential in food safety and environmental protection.

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基于多指示剂pH比色法和智能手机成像平台的便携式鼠伤寒沙门菌检测CRISPR/Cas12a生物传感器

传统的比色CRISPR/Cas方法依赖于单一的显色底物和笨重的专用信号检测仪器，这阻碍了它们的实际应用。本文首次报道了一种便携式和可访问的CRISPR/Cas12a生物传感器，该传感器利用多指示剂pH毫米盘（millidisc）进行信号可视化，并结合基于智能手机的成像平台进行信号读出，可对肠炎沙门氏菌血清型鼠伤寒沙门氏菌（S. typhimurium）进行敏感定量。pH微盘由多个指示剂组成，具有敏感的pH响应性，颜色变化多样。自主开发的RGB小程序名为DeepFood，专为便携式智能手机设计，具有用户友好的操作和趋势可视化，用于初步结果分析。根据鼠伤寒沙门氏菌RGB信号的变化规律，设计了一种独特的Senh信号类型，将信噪比从3.38提高到7.11。与R信号型相比，Senh信号型在0.01 M PBS缓冲液中检测灵敏度提高36.23倍（7.26 CFU/mL），在鸡中检测灵敏度提高15.53倍（1.41 × 102 CFU/mL）。该传感器在检测灵敏度和实用性方面有显著提高，在食品安全和环境保护方面具有潜力。

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

Biosensors and Bioelectronics 工程技术-电化学

CiteScore

20.80

自引率

7.10%

发文量

1006

审稿时长

29 days

期刊介绍： Biosensors & Bioelectronics, along with its open access companion journal Biosensors & Bioelectronics: X, is the leading international publication in the field of biosensors and bioelectronics. It covers research, design, development, and application of biosensors, which are analytical devices incorporating biological materials with physicochemical transducers. These devices, including sensors, DNA chips, electronic noses, and lab-on-a-chip, produce digital signals proportional to specific analytes. Examples include immunosensors and enzyme-based biosensors, applied in various fields such as medicine, environmental monitoring, and food industry. The journal also focuses on molecular and supramolecular structures for enhancing device performance.