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聚合物点用于在单个液滴中定量测定总疏水性致病裂解物。

Polymer dots for quantifying the total hydrophobic pathogenic lysates in a single drop.

机构信息

Department of Chemistry, National Sun Yat-Sen University, Kaohsiung 804, Taiwan; Department of Chemistry, Assuit University, Assuit 71516, Egypt.

Department of Chemistry, National Sun Yat-Sen University, Kaohsiung 804, Taiwan; School of Pharmacy, College of Pharmacy, Kaohsiung Medical University, Kaohsiung 806, Taiwan; Center for Nanoscience and Nanotechnology, National Sun Yat-Sen University, Kaohsiung 804, Taiwan; Doctoral Degree Program in Marine Biotechnology, National Sun Yat-Sen University, Kaohsiung 804, Taiwan.

出版信息

Colloids Surf B Biointerfaces. 2014 Mar 1;115:51-60. doi: 10.1016/j.colsurfb.2013.11.013. Epub 2013 Nov 20.

Abstract

A selective, rapid, small sample load (2-4 μL), and sensitive quantification method for the hydrophobic cellular biomolecules of pathogenic bacteria and their biosensing application were reported. The present approach is based on using polythiophene polymer dots (2.5 nm), which were prepared via the oxidation/polymerization reactions and then were characterized using transmission electron microscopy (TEM), Fourier transform infrared (FT-IR), ultraviolet spectroscopy (UV), and matrix (surface) assisted laser desorption/ionization mass spectrometry (M(S)ALDI-MS). The present method requires only gentle agitation for a single drop of aqueous bacteria suspension (10 μL, Pseudomonas aeruginosa (1×10(4) cfu/mL) and Staphylococcus aureus (1×10(5) cfu/mL)) with 1 mL of polythiophene (0.5 mg/mL) in chloroform, and the time required for quantifying the total hydrophobic was significantly reduced to less than 3 min. The polythiophene polymer dots is also a quantitative assessment of bacteria for aqueous and blood samples if exposed to more than 4-5 μL of pathogenic bacteria and thus, it is a new biosensor for quantitative hydrophobic portions. The fluorescence intensity of polythiophene was enhanced after adding different volumes of pathogenic bacteria with low colony units. The standard bacteria suspensions of P. aeruginosa and S. aureus have low LOD (limits of detection) for 2 μL (1×10(4) cfu/mL) and 4 μL (1×10(5) cfu/mL), respectively. Further, the pathogenic bacteria were spiked into mouse blood and the total hydrophobic biomolecules were quantified. This method is extremely rapid as it does not require any culture steps prior to analysis and also no need for any separation or post sample treatments.

摘要

报道了一种用于疏水性细胞生物分子的选择性、快速、小样本负载(2-4 μL)和灵敏定量方法及其生物传感应用。本方法基于使用聚噻吩聚合物点(2.5nm),通过氧化/聚合反应制备,然后使用透射电子显微镜(TEM)、傅里叶变换红外(FT-IR)、紫外光谱(UV)和基质(表面)辅助激光解吸/电离质谱(M(S)ALDI-MS)进行表征。本方法仅需轻轻搅拌单滴含 10μL 水的细菌悬浮液(1×10(4) cfu/mL 铜绿假单胞菌和 1×10(5) cfu/mL 金黄色葡萄球菌)和 1 mL 浓度为 0.5mg/mL 的氯仿中的聚噻吩聚合物点,定量总疏水性所需的时间显著缩短至 3 分钟以内。如果暴露于超过 4-5μL 的病原菌,聚噻吩聚合物点还可以定量评估水相和血样中的细菌,因此,它是一种新的定量疏水部分生物传感器。添加不同体积的低菌落单位病原菌后,聚噻吩的荧光强度增强。P. aeruginosa 和 S. aureus 的标准细菌悬浮液的检测限(LOD)均为 2μL(1×10(4) cfu/mL)和 4μL(1×10(5) cfu/mL)。此外,将病原菌掺入小鼠血液中并定量总疏水性生物分子。该方法非常快速,因为在分析之前不需要任何培养步骤,也不需要任何分离或样品后处理。

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