■ε毒素(ETX),产气荚膜梭菌,是已知最有效的毒素之一,具有接近肉毒杆菌神经毒素的致命效力。ε毒素是肠炎的原因。因此,发展快速简便的ETX检测方法势在必行。适体是单链寡核苷酸,其可以以与单克隆抗体(mAb)相当的亲和力与特异性靶分子紧密结合。DNA适体可以作为生物分子鉴定的工具,如病原体亚种。
■本研究旨在分离抗ETX的高亲和力单链DNA(ssDNA)适体。
■这项研究使用指数富集(SELEX)方法对配体进行系统进化,酶联apta吸附剂测定(ELASA),和表面等离子体共振(SPR)来确定新获得的靶向ETX的适配体的亲和力和特异性。
■研究了通过SELEX方法获得的几种适体。其中,2个适体,ETX克隆3(ETX3;解离常数(Kd=8.4±2.4E-9M)和ETX11(Kd=6.3±1.3E-9M)对ETX具有良好的特异性。ETX3和ETX11的检出限分别为0.21和0.08μg/mL。.
■发现的适体可用于各种基于适体的快速诊断测试中,以检测ETX。
UNASSIGNED: Epsilon toxin (ETX), produced by Clostridium perfringens, is one of the most potent toxins known, with a lethal potency approaching that of botulinum neurotoxins. Epsilon toxin is responsible for enteritis. Therefore, the development of rapid and simple methods to detect ETX is imperative. Aptamers are single-stranded oligonucleotides that can bind tightly to specific target molecules with an affinity comparable to that of monoclonal antibodies (mAbs). DNA aptamers can serve as tools for the molecular identification of organisms, such as pathogen subspecies.
UNASSIGNED: This study aimed to isolate high-affinity single-stranded DNA (ssDNA) aptamers against ETX.
UNASSIGNED: This study identified aptamers using the Systematic Evolution of Ligands by Exponential Enrichment (SELEX) method, enzyme-linked apta-sorbent assay (
ELASA), and surface plasmon resonance (SPR) to determine the affinity and specificity of the newly obtained aptamers targeting ETX.
UNASSIGNED: Several aptamers obtained through the SELEX process were studied. Among them, 2 aptamers, ETX clone 3 (ETX3; dissociation constant (Kd = 8.4 ± 2.4E-9M) and ETX11 (Kd = 6.3 ± 1.3E-9M) had favorable specificity for ETX. The limits of detection were 0.21 and 0.08 μg/mL for ETX3 and ETX11, respectively..
UNASSIGNED: The discovered aptamers can be used in various aptamer-based rapid diagnostic tests for the detection of ETX.