Instituto de Medicina Tropical e Departamento de Moléstias Infecciosas e Parasitarias da Faculdade de Medicina da Universidade de São Paulo, São Paulo, Brazil.
Biologia Molecular, Clinimol Diag Molecular, Sao Paulo, Brazil.
Front Cell Infect Microbiol. 2022 Jun 30;12:818737. doi: 10.3389/fcimb.2022.818737. eCollection 2022.
Infections caused by multidrug-resistant are a worldwide health concern with high mortality rates. Rapid identification of this infectious agent is critical as it can easily spread with difficult or no options for treatment. In this context, the development of reliable and economically viable detection and therapeutic methodologies are still challenging. One of the promising solutions is the development of nucleic acid aptamers capable of interacting with bacteria. These aptamers can be used for specific recognition of infectious agents as well as for blocking their functions. Cell-SELEX technology currently allows the selection and identification of aptamers and is flexible enough to target molecules present in an entire bacterial cell without their prior knowledge. However, the aptamer technology is still facing many challenges, such as the complexity of the screening process. Here, we describe the selection and identification of a new aptamer A01, using an in-house whole-cell SELEX-based methodology, against multi-resistant , with rapid execution and low cost. In addition, this protocol allowed the identification of the aptamer A01 with the whole cell as a target. The aptamer A01 demonstrated a binding preference to when compared to , , and in fluorescence assays. Although the time-kill assay did not show an effect on bacterial growth, the potential bactericidal or bacteriostatic cannot be totally discarded. The new categorized aptamer (A01) displayed a significant binding affinity to MDR .
由耐多药引起的感染是一个全球性的健康问题，死亡率很高。快速识别这种感染因子至关重要，因为它很容易传播，而且治疗选择困难或没有。在这种情况下，开发可靠且经济可行的检测和治疗方法仍然具有挑战性。有希望的解决方案之一是开发能够与细菌相互作用的核酸适体。这些适体可用于特异性识别感染因子，并阻断其功能。细胞 SELEX 技术目前允许选择和鉴定适体，并且足够灵活，可以针对整个细菌细胞中存在的分子进行靶向，而无需事先了解这些分子。然而，适体技术仍然面临许多挑战，例如筛选过程的复杂性。在这里，我们描述了使用内部全细胞 SELEX 为基础的方法对多药耐药进行了新的适体 A01 的选择和鉴定，该方法执行速度快，成本低。此外，该方案允许将适体 A01 与整个细胞作为靶标进行鉴定。与 、 和 相比，荧光分析显示适体 A01 对 具有结合偏好。虽然时间杀伤试验并未显示对细菌生长有影响，但不能完全排除潜在的杀菌或抑菌作用。新型分类适体（A01）对 MDR 表现出显著的结合亲和力。
