Breaking barrier of binding buffer in colorimetric aptasensing of tetracycline in food fish using peroxidase mimic gold NanoZyme

IF 4.2 3区生物学 Q1 BIOCHEMICAL RESEARCH METHODS

Methods Pub Date : 2025-04-28 DOI:10.1016/j.ymeth.2025.04.017

Dhruba Jyoti Sarkar , Ramij Raja , V. Santhana Kumar , Soumyadeb Bhattacharyya , Souvik Pal , Subhankar Mukherjee , Basanta Kumar Das

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Abstract

Tetracycline is extensively used in aquaculture as a therapeutic agent that needs to be monitored due to food safety concerns. Aptasensing has been revealed as a suitable diagnostic platform for tetracycline sensing in food matrix due to its quick, low cost and robust nature. But, the colorimetric aptasensing of tetracycline employing the peroxidase activity of gold nanoparticles (AuNPs) to 3,3,5,5-tetramethylbenzidine (TMB) was unsuitable until now owing to the aptamer-specific alkaline binding buffer. The present study developed a method with an optimized reaction protocol diminishing the inhibitory effect of binding buffer on the sensor probe (AuNPs-aptamer + TMB + H₂O₂). The overall peroxidase activity of the sensor probe was only inhibited by tetracycline through selective adsorption on the AuNPs-aptamer complex. The peroxidase inhibition percentage in the test range of 0.01 to 0.5 mg L^-1 tetracycline gave a logarithmic response (R², 0.99) with a detection limit of 0.017 mg L^-1 which is less than the prescribed limit (0.1 mg L^-1) set by EU and FSSAI. The developed sensing system in fish muscle showed high recovery (111–115 %) with great potential for rapid detection of tetracycline in fish muscle.

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利用过氧化物酶模拟金纳米酶对食用鱼中四环素进行比色感应，突破结合缓冲液的屏障

四环素作为一种治疗剂广泛用于水产养殖，由于食品安全问题，需要对其进行监测。Aptasensing因其快速、低成本和鲁棒性而被认为是一种适合于食品基质中四环素检测的诊断平台。但是，利用金纳米粒子（AuNPs）的过氧化物酶活性对3,3,5,5-四甲基联苯胺（TMB）进行比色感应四环素，由于适配体特异性碱性结合缓冲液的存在，目前尚不适合。本研究开发了一种优化反应方案的方法，减少了结合缓冲液对传感器探针（AuNPs-aptamer + TMB + H2O2）的抑制作用。传感器探针的总过氧化物酶活性仅通过四环素在aunps -适配体复合物上的选择性吸附被抑制。四环素对过氧化物酶的抑制率在0.01 ~ 0.5 mg L-1范围内呈对数响应（R2 = 0.99），检出限为0.017 mg L-1，低于欧盟和FSSAI规定的检测限（0.1 mg L-1）。该检测系统回收率高（111 ~ 115%），具有快速检测鱼类肌肉中四环素的潜力。

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来源期刊

Methods 生物-生化研究方法

CiteScore

9.80

自引率

2.10%

发文量

222

审稿时长

11.3 weeks

期刊介绍： Methods focuses on rapidly developing techniques in the experimental biological and medical sciences. Each topical issue, organized by a guest editor who is an expert in the area covered, consists solely of invited quality articles by specialist authors, many of them reviews. Issues are devoted to specific technical approaches with emphasis on clear detailed descriptions of protocols that allow them to be reproduced easily. The background information provided enables researchers to understand the principles underlying the methods; other helpful sections include comparisons of alternative methods giving the advantages and disadvantages of particular methods, guidance on avoiding potential pitfalls, and suggestions for troubleshooting.