A Polyclonal Aptamer Library for the Specific Binding of the Gut Bacterium Roseburia intestinalis in Mixtures with Other Gut Microbiome Bacteria and Human Stool Samples

Roseburia intestinalis has received attention as a potential probiotic bacterium. Recent studies have demonstrated that changes in its intestinal abundance can cause various diseases, such as obesity, enteritis and atherosclerosis. Probiotic administration or fecal transplantation alter the structure of the intestinal flora, offering possibilities for the prevention and treatment of these diseases. However, current monitoring methods, such as 16S rRNA sequencing, are complex and costly and require specialized personnel to perform the tests, making it difficult to continuously monitor patients during treatment. Hence, the rapid and cost-effective quantification of intestinal bacteria has become an urgent problem to be solved. Aptamers are of emerging interest because their stability, low immunogenicity and ease of modification are attractive properties for a variety of applications. We report a FluCell-SELEX polyclonal aptamer library specific for R. intestinalis isolated after seven evolution rounds, that can bind and label this organism for fluorescence microscopy and binding assays. Moreover, R. intestinalis can be distinguished from other major intestinal bacteria in complex defined mixtures and in human stool samples. We believe that this preliminary evidence opens new avenues towards aptamer-based electronic biosensors as new powerful and inexpensive diagnostic tools for the relative quantitative monitoring of R. intestinalis in gut microbiomes.     

Relative Nuclease Resistance of a DNA Aptamer Covalently Conjugated to a Target Protein

A major obstacle to the therapeutic application of an aptamer is its susceptibility to nuclease digestion. Here, we confirmed the acquisition of relative nuclease resistance of a DNA-type thrombin binding aptamer with a warhead (TBA₃) by covalent binding to a target protein in the presence of serum/various nucleases. When the thrombin-inhibitory activity of TBA₃ on thrombin was reversed by the addition of the complementary strand, the aptamer was instantly degraded by the nucleases, showing that the properly folded/bound aptamer conferred the resistance. Covalently binding aptamers possessing both a prolonged drug effect and relative nuclease resistance would be beneficial for in vivo translational applications.     

Unmodified gold nanoparticles as a colorimetric probe for visual methamphetamine detection

In recent years, in addition to the classic drugs, addiction to a series of new drug classes known as club drugs has increased significantly. Fast and low-cost bioassay for the detection of amphetamine-based drugs can be an effective strategy towards reducing their abuse. In this study, we designed a sensitive bioassay strategy using gold nanoparticles (GNPs) and the aptamers that possess high affinity toward methamphetamine (MA). It is suggested that the aptamer adopts different tertiary structures in the presence and/or absence of its specific target and GNPs can effectively differentiate between these two states by their characteristic surface plasmon resonance-based colour change. Visual detection of MA and 3,4-methylenedioxy-N-methylamphetamine (MDMA) in the low micromolar range is possible within minutes with the use of this method.     

基于人工模拟抗体的酶联检测技术在食品安全检测中的应用

广义人工模拟抗体包括适配体、重组受体、分子印迹聚合物及多肽。人工模拟抗体与传统抗体相比优势显著,其具有易实现工业化生产、成本低、保存方便、可反复使用等特点。酶联检测技术是人工模拟抗体在食品安全检测中应用最为广泛的技术。本文综述了人工模拟抗体的特点以及基于人工模拟抗体的酶联检测技术在食品安全检测中的应用。     

甲胺磷的核酸适体筛选

目的 筛选识别甲胺磷的DNA适体。方法 体外合成ssDNA 文库, 将其固相化于琼脂糖凝胶颗粒表面, 液相中甲胺磷与其结合的DNA分子从固相表面分离, 进而建立非固相化SELEX技术, 最终获得识别甲胺磷的适体, 采用DNAMAN和RNA structure软件对适体进行一、二级结构分析。结果 经过10 轮筛选, ssDNA 文库与甲胺磷的亲和力呈上升趋势, 随机挑选的20个阳性克隆适体根据一级结构的同源性可分为5个家族, 二级结构预测以茎环结构为主。结论 本方法大大提高了适体的筛选效率, 最终筛选得到能识别甲胺磷的DNA适体。     

基于纳米金与适配体的赭曲霉毒素A检测方法研究

以赭曲霉毒素A适配体作为识别分子荧光素发射的荧光作为检测信号、纳米金作为荧光信号淬灭剂,建立一种高灵敏度的赭曲霉毒素A荧光检测方法。结果表明:该检测体系检测限达到10nmol/L,在25~1 000nmol/L浓度范围具有线性关系,相关系数为0.991,具有较好的实用性。该荧光检测法同传统方法相比,提高了检测效率,为更好地进行赭曲霉毒素A检测控制提供了技术支持。     

乳制品中功能性蛋白适配体传感器的构建及其在检测中的应用

乳制品中功能性蛋白包括乳铁蛋白、乳桥蛋白和β-乳球蛋白等,具有抗微生物、抗菌和调节免疫系统等生理功能;同时,β-乳球蛋白、α-乳清蛋白和酪蛋白等会引起部分人群发生过敏反应。因此,定量检测乳制品中功能性蛋白对其品质评价至关重要。适配体传感器技术是一种高特异性、高灵敏度的快速检测方法,目前研究者们通过多种筛选方法获得功能性蛋白的适配体,并构建了用于功能性蛋白检测的适配体传感器。本文综述了近年来针对乳制品中功能性蛋白的适配体检测技术的研究进展,重点介绍了乳铁蛋白、乳桥蛋白、β-乳球蛋白、α-乳清蛋白和酪蛋白的适配体筛选技术及其适配体传感器的构建与应用,为进一步开发便捷、灵敏、高效的生物传感器及分析方法提供参考。     

仿生识别传感器的制备及其在抗生素检测中的应用

基于仿生识别的生物传感器作为一种新兴的检测手段,具有高效、低成本、灵敏度高和特异性强等显著优势,非常适合于复杂基质样品中痕量目标物的快速检测。为更好地研究开发新的生物传感器,该文对近几年来几种常见生物传感器进行分类综述,着重分析主要仿生识别受体和换能器的基本原理及其优缺点,以及它们在食品抗生素残留检测中的应用。最后,对仿生识别手段的发展前景进行展望。