利用aptamer横向流动检测法优化鼠伤寒沙门氏菌的检测。

IF 2 4区 生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Biotechnology Letters Pub Date : 2024-08-01 Epub Date: 2024-05-28 DOI:10.1007/s10529-024-03484-1
Nafise Abedi, Mehdi Zeinoddini, Mohammad Shoushtari
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引用次数: 0

摘要

伤寒沙门氏菌是一种对医学和食品工业有重大影响的致病菌,它可引起伤寒等食源性疾病,对人类构成严重威胁。由于伤寒杆菌的症状与多种疾病重叠,因此准确诊断伤寒杆菌具有挑战性。因此,我们需要一种精确、高效的诊断方法。在本研究中,我们采用田口优化法开发了一种基于适配体侧流分析法(LFA)的生物传感器,用于检测伤寒杆菌。因此,我们使用抗沙门氏菌适配体设计了信号探针和纳米生物探针,并与金纳米粒子(GNPs)共轭,用于 LFA。该检测的策略基于固定在膜上的细菌与检测样品中存在的细菌之间的竞争形式。此外,还采用田口试验设计法对影响适配体 LFA 的各种因素进行了优化,包括细菌的浓度(固定细菌和样品中的细菌)和纳米生物胶的浓度。数据显示,LFA 反应的最佳条件是固定化细菌浓度为 108 CFU/mL,纳米杀菌剂浓度为 1.5 μg/μL。然后,伤寒杆菌的肉眼检测限估计为 105 CFU/mL。反应结果在 20 分钟内得出,与其他食品病原体没有明显的交叉反应。总之,采用田口方法优化的适配体-LFA 诊断方法是检测鼠伤寒杆菌的一种可靠、直接和准确的工具。总之,该方法可作为检测和鉴定细菌的便携式诊断试剂盒。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Optimized detection of Salmonella typhimurium using aptamer lateral flow assay.

Optimized detection of Salmonella typhimurium using aptamer lateral flow assay.

Salmonella typhimurium, a pathogenic bacterium with significant implications in medicine and the food industry, poses a substantial threat by causing foodborne illnesses such as typhoid fever. Accurate diagnosis of S. typhimurium is challenging due to its overlap symptoms with various diseases. This underscores the need for a precise and efficient diagnostic approach. In this study, we developed a biosensor using the Taguchi optimization method based on aptamer lateral flow assay (LFA) for the detection of S. typhimurium. Therefore, signal probe and nanobioprobe were designed using anti-Salmonella aptamer, conjugated with gold nanoparticles (GNPs), and used in LFA. The strategy of this test is based on a competitive format between the bacteria immobilized on the membrane and the bacteria present in the tested sample. Moreovere, the optimization of various factors affecting the aptamer LFA, including the concentration of bacteria (immobilized and into the sample) and the concentration of nanobioprop, were performed using the Taguchi test designing method. The data showed that the optimal conditions for the LFA reaction was 108 CFU/mL of immobilized bacteria and 1.5 μg/μL of nanobioprop concentration. Then, the visual detection limit of S. typhimurium was estimated as 105 CFU/mL. The reaction results were obtained within 20 min, and there were no significant cross-reactions with other food pathogens. In conclusion, the aptamer-LFA diagnostic method, optimized using the Taguchi approach, emerges as a reliable, straightforward, and accurate tool for the detection of S. typhimurium. Overall, this method can be a portable diagnostic kit for the detection and identification of bacteria.

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来源期刊
Biotechnology Letters
Biotechnology Letters 工程技术-生物工程与应用微生物
CiteScore
5.90
自引率
3.70%
发文量
108
审稿时长
1.2 months
期刊介绍: Biotechnology Letters is the world’s leading rapid-publication primary journal dedicated to biotechnology as a whole – that is to topics relating to actual or potential applications of biological reactions affected by microbial, plant or animal cells and biocatalysts derived from them. All relevant aspects of molecular biology, genetics and cell biochemistry, of process and reactor design, of pre- and post-treatment steps, and of manufacturing or service operations are therefore included. Contributions from industrial and academic laboratories are equally welcome. We also welcome contributions covering biotechnological aspects of regenerative medicine and biomaterials and also cancer biotechnology. Criteria for the acceptance of papers relate to our aim of publishing useful and informative results that will be of value to other workers in related fields. The emphasis is very much on novelty and immediacy in order to justify rapid publication of authors’ results. It should be noted, however, that we do not normally publish papers (but this is not absolute) that deal with unidentified consortia of microorganisms (e.g. as in activated sludge) as these results may not be easily reproducible in other laboratories. Papers describing the isolation and identification of microorganisms are not regarded as appropriate but such information can be appended as supporting information to a paper. Papers dealing with simple process development are usually considered to lack sufficient novelty or interest to warrant publication.
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