Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.
Applied Microbiology Research Center, Biomedicine Technologies Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran.
Mol Biol Rep. 2024 Jun 3;51(1):722. doi: 10.1007/s11033-024-09561-y.
Yersinia pestis is a bacterium that causes the disease plague. It has caused the deaths of many people throughout history. The bacterium possesses several virulence factors (pPla, pFra, and PYV). PFra plasmid encodes fraction 1 (F1) capsular antigen. F1 protein protects the bacterium against host immune cells through phagocytosis process. This protein is specific for Y. pestis. Many diagnostic techniques are based on molecular and serological detection and quantification of F1 protein in different food and clinical samples. Aptamers are small nucleic acid sequences that can act as specific ligands for many targets.This study, aimed to isolate the high-affinity ssDNA aptamers against F1 protein.
In this study, SELEX was used as the main strategy in screening aptamers. Moreover, enzyme-linked aptamer sorbent assay (ELASA) and surface plasmon resonance (SPR) were used to determine the affinity and specificity of obtained aptamers to F1 protein. The analysis showed that among the obtained aptamers, the three aptamers of Yer 21, Yer 24, and Yer 25 were selected with a KD value of 1.344E - 7, 2.004E - 8, and 1.68E - 8 M, respectively. The limit of detection (LoD) was found to be 0.05, 0.076, and 0.033 μg/ml for Yer 21, Yer 24, and Yer 25, respectively.
This study demonstrated that the synthesized aptamers could serve as effective tools for detecting and analyzing the F1 protein, indicating their potential value in future diagnostic applications.
鼠疫耶尔森菌是一种导致鼠疫的细菌，它在历史上导致了许多人的死亡。该细菌拥有几种毒力因子（pPla、pFra 和 PYV）。pFra 质粒编码分 1（F1）荚膜抗原。F1 蛋白通过吞噬作用保护细菌免受宿主免疫细胞的攻击。这种蛋白质是鼠疫耶尔森菌特有的。许多诊断技术基于分子和血清学检测以及不同食品和临床样本中 F1 蛋白的定量。适体是可以作为许多靶标特异性配体的小核酸序列。本研究旨在分离针对 F1 蛋白的高亲和力 ssDNA 适体。
在这项研究中，SELEX 被用作筛选适体的主要策略。此外，酶联适体吸附测定（ELASA）和表面等离子体共振（SPR）用于确定获得的适体与 F1 蛋白的亲和力和特异性。分析表明，在所获得的适体中，Yer 21、Yer 24 和 Yer 25 三个适体的 KD 值分别为 1.344E-7、2.004E-8 和 1.68E-8M。发现 Yer 21、Yer 24 和 Yer 25 的检测限（LoD）分别为 0.05、0.076 和 0.033μg/ml。
本研究表明，合成的适体可以作为检测和分析 F1 蛋白的有效工具，表明它们在未来的诊断应用中具有潜在价值。
