用 PGM 进行铜绿假单胞菌的 POCT 检测及在预防支气管镜检查中的非医院感染中的应用。

IF 2.3 3区 化学 Q3 CHEMISTRY, ANALYTICAL Journal of Analytical Methods in Chemistry Pub Date : 2024-09-05 DOI:10.1155/2024/8062001
Tao Wen,Houqi Ning,Yinping Yang,Jinze Zhang
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引用次数: 0

摘要

背景造成支气管镜污染的主要病原体是铜绿假单胞菌。传统的支气管镜消毒技术和病原体鉴定方法具有耗时长、操作复杂等特点。本研究的目的是建立一种快速、精确的铜绿假单胞菌鉴定方法,最终目标是降低与该病原体相关的院内感染风险。方法先一步合成磁性纳米粒子(MNPs),然后用抗体和转化酶优化涂层工艺,生产出双功能化的 IMIc。单克隆抗体被固定在微孔板上,用于特异性捕获和富集铜绿假单胞菌。铜绿假单胞菌存在时,单克隆抗体、测试样本和 IMIc 形成夹层结构。随后加入蔗糖溶液,通过个人葡萄糖计检测转化酶水解产生的葡萄糖,从而定量评估铜绿假单胞菌的浓度。NTA 确定磁性纳米粒子的晶粒直径为 200 nm。傅立叶变换红外光谱显示,MNPs 上的两个羧基被成功修饰。优化微孔板包被抗体的孵育 pH 值为 7;优化微孔板包被抗体的孵育时间为 2 小时;优化多克隆抗体的连接 pH 值为 5;与磁珠连接的多克隆抗体的反应时间为 1 小时;与磁珠连接的转化酶的 pH 值为 4。PGM 是一种方便、准确的定量检测方法,适用于潜在的临床诊断应用。
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POCT Detection of Pseudomonas aeruginosa by PGM and Application of Preventing Nosocomial Infection of Bronchoscopy.
Background The primary pathogen responsible for bronchoscope contamination is Pseudomonas aeruginosa. Conventional techniques for bronchoscopy disinfection and pathogen identification methods are characterized by time-consuming and operation complexly. The objective of this research is to establish a prompt and precise method for the identification of Pseudomonas aeruginosa, with the ultimate goal of mitigating the risk of nosocomial infections linked to this pathogen. Methods The magnetic nanoparticles (MNPs) were synthesized in a single step, followed by the optimization of the coating process with antibodies and invertase to produce the bifunctionalized IMIc. Monoclonal antibodies were immobilized on microplates for the specific capture and enrichment of Pseudomonas aeruginosa. Upon the presence of Pseudomonas aeruginosa, the monoclonal antibodies, the test sample, and the IMIc formed sandwich structures. The subsequent addition of a sucrose solution allowed for the detection of glucose produced through invertase hydrolysis by a personal glucose meter, enabling quantitative assessment of Pseudomonas aeruginosa concentration. Results TEM image demonstrates that the MNPs exhibit a consistent spherical shape. NTA determined that the grain diameter of magnetic nanoparticles was 200 nm. FTIR spectrum revealed the successful modification of two carboxyl groups on the MNPs. The optimization of the incubation pH of the microplate-coated antibody was 7. The optimization of the incubation time of the microplate-coated antibody was 2 h. The optimization of the ligation pH for the polyclonal antibody was 5. Reaction times of polyclonal antibodies linked to magnetic beads was 1 h. The pH of invertase linked by magnetic beads was 4. Conclusion This article presents a novel qualitative and quantitative immunoassay for point-of-care monitoring of P. aeruginosa utilizing PGM as a readout. The PGM represents a convenient and accurate quantitative detection method suitable for potential clinical diagnostic applications.
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来源期刊
Journal of Analytical Methods in Chemistry
Journal of Analytical Methods in Chemistry CHEMISTRY, ANALYTICAL-ENGINEERING, CIVIL
CiteScore
4.80
自引率
3.80%
发文量
79
审稿时长
6-12 weeks
期刊介绍: Journal of Analytical Methods in Chemistry publishes papers reporting methods and instrumentation for chemical analysis, and their application to real-world problems. Articles may be either practical or theoretical. Subject areas include (but are by no means limited to): Separation Spectroscopy Mass spectrometry Chromatography Analytical Sample Preparation Electrochemical analysis Hyphenated techniques Data processing As well as original research, Journal of Analytical Methods in Chemistry also publishes focused review articles that examine the state of the art, identify emerging trends, and suggest future directions for developing fields.
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