State Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi 214122, China.
J Agric Food Chem. 2012 Apr 25;60(16):4034-8. doi: 10.1021/jf300395z. Epub 2012 Apr 13.
A whole-bacterium systemic evolution of ligands by exponential enrichment (SELEX) method was applied to a combinatorial library of FAM-labeled single-stranded DNA molecules to identify DNA aptamers demonstrating specific binding to Vibrio parahemolyticus . FAM-labeled aptamer sequences with high binding affinity to V. parahemolyticus were identified by flow cytometric analysis. Aptamer A3P, which showed a particularly high binding affinity in preliminary studies, was chosen for further characterization. This aptamer displayed a dissociation constant (K(d)) of 16.88 ± 1.92 nM. Binding assays to assess the specificity of aptamer A3P showed a high binding affinity (76%) for V. parahemolyticus and a low apparent binding affinity (4%) for other bacteria. Whole-bacterium SELEX is a promising technique for the design of aptamer-based molecular probes for microbial pathogens that does not require the labor-intensive steps of isolating and purifying complex markers or targets.
采用全菌体配体指数富集系统进化（SELEX）方法，对 FAM 标记的单链 DNA 分子组合文库进行筛选，以鉴定与副溶血性弧菌具有特异性结合的 DNA 适体。通过流式细胞术分析鉴定出与副溶血性弧菌具有高结合亲和力的 FAM 标记适体序列。在初步研究中表现出特别高结合亲和力的适体 A3P 被选择用于进一步表征。该适体的解离常数（Kd）为 16.88 ± 1.92 nM。为评估适体 A3P 的特异性而进行的结合测定显示，该适体对副溶血性弧菌具有高结合亲和力（76%），对其他细菌的表观结合亲和力较低（4%）。全菌体 SELEX 是一种很有前途的技术，可用于设计基于适体的微生物病原体分子探针，该技术不需要进行费力的分离和纯化复杂标记物或靶标的步骤。
