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通过 Cas12a-RAA 与 LFS/PGM 双信号读出生物传感器相结合,实现对动物源食品中沙门氏菌的便携检测。

Portable detection of Salmonella in food of animal origin via Cas12a-RAA combined with an LFS/PGM dual-signaling readout biosensor.

机构信息

Institute of Food & Nutrition Science and Technology, Shandong Provincial Key Laboratory of Agro-Products Processing Technology, Key Laboratory of Novel Food Resources Processing, Ministry of Agriculture, Shandong Academy of Agricultural Sciences, Jinan, 250100, China.

College of Food Science and Technology, Huazhong Agricultural University, Wuhan, 430070, China.

出版信息

Mikrochim Acta. 2024 Sep 28;191(10):631. doi: 10.1007/s00604-024-06708-3.

Abstract

A highly specific and sensitive rapid two-signal assay was developed for the detection of Salmonella typhimurium in foods of animal origin. The invA gene of Salmonella was used as the biorecognition element and recombinase-assisted amplification (RAA) technology for signal amplification. By utilizing the specific recognition and efficient trans-cleavage activity of CRISPR/Cas12a, point-of-care testing (POCT) for S. typhimurium was achieved via lateral flow strips (LFS) and personal glucometer (PGM) biosensors as dual signal readout systems, with sensitivities of 33 CFU/mL and 20 CFU/mL, respectively. Users can select the appropriate test system on the basis of specific application requirements: LFSs are ideal for rapid onsite screening, whereas glucometer biosensors offer precise quantitative determination. This approach simplifies the use of large instruments and overcomes site constraints, demonstrating good accuracy and applicability in animal-derived samples, with significant potential for the detection of other pathogens and for use in restricted environments.

摘要

本研究开发了一种用于检测动物源食品中鼠伤寒沙门氏菌的高特异性和高灵敏度的双信号快速检测方法。该方法以沙门氏菌的 invA 基因为生物识别元件,重组酶辅助扩增(RAA)技术为信号放大技术。通过利用 CRISPR/Cas12a 的特异性识别和高效的转切割活性,基于横向流条(LFS)和个人血糖仪(PGM)生物传感器的即时检测(POCT)可作为双信号读出系统,检测鼠伤寒沙门氏菌的灵敏度分别为 33 CFU/mL 和 20 CFU/mL。用户可以根据特定的应用要求选择合适的测试系统:LFS 非常适合快速现场筛查,而血糖仪生物传感器则可提供精确的定量测定。这种方法简化了大型仪器的使用,克服了现场限制,在动物源性样本中具有良好的准确性和适用性,在检测其他病原体和在受限环境中使用方面具有很大的潜力。

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