Novel strategy for screening aptamers of Staphylococcus aureus enterotoxin A based on active fragments and fusion design

IF 5.7 2区化学 Q1 CHEMISTRY, ANALYTICAL

Analytica Chimica Acta Pub Date : 2025-04-30 DOI:10.1016/j.aca.2025.344135

Shujing Ge, Mengyao Guan, Fei Pan, Yanli Wang, Daodong Pan, Chaoying Zhang, Xinchang Gao, Yali Dang

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引用次数: 0

Abstract

Background

Staphylococcus aureus enterotoxin A (SEA) is the major toxin responsible for food poisoning caused by Staphylococcus aureus infection. Aptamers are an ideal solution for visual detection of SEA. However, screening for aptamers is generally expensive and time-consuming. Therefore, a new method for efficient screening of high-affinity SEA aptamers needs to be established and supplemented with smartphone image recognition to enable visual and rapid detection of SEA.

Results

A new aptamer for sensitive detection of SEA was screened using active fragments and fusion design. The novel high-affinity aptamer (BX1) was derived by intercepting active nucleotide fragments of several aptamers and fusing them; it was verified to have high affinity, with a K_d value of 4.16 ± 0.22 nM. Molecular dynamic simulation illustrated that BX1 formed a stable complex conformation with SEA, which maintained its stability for 200 ns. This novel aptamer can be used for visual inspection of SEA. A smartphone-assisted aptasensor was designed to sensitively detect SEA in milk with a wide linear range (1-250 ng/mL), low detection limit (0.42 ng/mL), and satisfactory spiked recovery.

Significance

A new method for screening SEA aptamers based on active fragments and fusion design was developed, which can rapidly screen high-affinity aptamers in a short duration and at a low expenditure. In addition, a new strategy for the rapid detection of SEA using a smartphone-assisted colorimetric aptamer sensor was successfully applied. This research method may provide a reference for the highly sensitive detection of other biotoxins.

Abstract Image

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基于活性片段和融合设计筛选金黄色葡萄球菌肠毒素A适体的新策略

背景金黄色葡萄球菌肠毒素A （SEA）是金黄色葡萄球菌感染引起食物中毒的主要毒素。适配体是SEA视觉检测的理想解决方案。然而，筛选适体通常既昂贵又耗时。因此，需要建立一种高效筛选高亲和力SEA适配体的新方法，并辅以智能手机图像识别，实现SEA的视觉快速检测。结果采用活性片段和融合设计筛选了一种新的SEA敏感检测适体。该新型高亲和力适配体（BX1）是通过截取多个适配体的活性核苷酸片段并融合得到的；结果表明，该蛋白具有较高的亲和力，Kd值为4.16±0.22 nM。分子动力学模拟表明，BX1与SEA形成稳定的络合构象，并在200 ns内保持稳定。该新型适配体可用于SEA的目视检测。该传感器线性范围宽（1 ~ 250 ng/mL），检出限低（0.42 ng/mL），加标回收率高，灵敏度高。研究了一种基于活性片段和融合设计的SEA核酸适体筛选新方法，该方法可以在短时间内以低成本快速筛选高亲和力的核酸适体。此外，还成功应用了一种利用智能手机辅助比色适体传感器快速检测SEA的新策略。该研究方法可为其他生物毒素的高灵敏度检测提供参考。

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来源期刊

Analytica Chimica Acta 化学-分析化学

CiteScore

10.40

自引率

6.50%

发文量

1081

审稿时长

38 days

期刊介绍： Analytica Chimica Acta has an open access mirror journal Analytica Chimica Acta: X, sharing the same aims and scope, editorial team, submission system and rigorous peer review. Analytica Chimica Acta provides a forum for the rapid publication of original research, and critical, comprehensive reviews dealing with all aspects of fundamental and applied modern analytical chemistry. The journal welcomes the submission of research papers which report studies concerning the development of new and significant analytical methodologies. In determining the suitability of submitted articles for publication, particular scrutiny will be placed on the degree of novelty and impact of the research and the extent to which it adds to the existing body of knowledge in analytical chemistry.