Integration of logic gates to CRISPR/Cas12a system for rapid and sensitive detection of pathogenic bacterial genes,Analytica Chimica Acta

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Integration of logic gates to CRISPR/Cas12a system for rapid and sensitive detection of pathogenic bacterial genes
Analytica Chimica Acta ( IF 5.7 ) Pub Date : 2020-08-01 , DOI: 10.1016/j.aca.2020.05.017
Lei Peng ₁ , Jin Zhou ₁ , Lijuan Yin ₁ , Shuli Man ₁ , Long Ma ₁

Affiliation

For the first time, three 2-input elementary AND, OR, INHIBIT logic gates have been constructed by using CRISPR-Cas12a system. These logic gates utilised the intrinsic advantages of programmability, sequence specificity and high base resolution of CRISPR-Cas12a system. Among them, the AND gate owned the potentials as a built-in biosensor that responded rapidly to external pathogenic bacteria such as Staphylococcus aureus with high sensitivity and specificity. We applied the CRISPR-Cas12a based bacterial detection after a target-amplification using PCR. The total sample-to-answer time was appropriately 2.0 h, the limit of detection (LOD) was 103 CFU/mL, and the dynamic range was 103-107 CFU/mL. Also, the sequence addressability enabled this AND logic gate to accurately trace back and distinguish input genes. These above-mentioned features were highly ideal to incur a rapid response to pathogenic bacteria for decision making. Our results not only validated the possibility of using CRISPR-Cas systems for constructing bio-computing devices but also provided a prototype of biosensor for rapid and intelligent pathogenic bacteria detection.

中文翻译：

将逻辑门集成到 CRISPR/Cas12a 系统以快速灵敏地检测病原细菌基因

首次使用CRISPR-Cas12a系统构建了三个2输入初等AND、OR、INHIBIT逻辑门。这些逻辑门利用了 CRISPR-Cas12a 系统的可编程性、序列特异性和高碱基分辨率的内在优势。其中，与门具有作为内置生物传感器的潜力，可以对金黄色葡萄球菌等外部病原菌快速反应，具有高灵敏度和特异性。我们在使用 PCR 进行目标扩增后应用基于 CRISPR-Cas12a 的细菌检测。总样本到回答时间为 2.0 小时，检测限 (LOD) 为 103 CFU/mL，动态范围为 103-107 CFU/mL。此外，序列可寻址性使该 AND 逻辑门能够准确追溯和区分输入基因。上述这些特征非常适合对病原菌做出快速反应以进行决策。我们的研究结果不仅验证了使用 CRISPR-Cas 系统构建生物计算设备的可能性，而且为快速智能检测病原菌提供了生物传感器的原型。

更新日期：2020-08-01

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