Abstract：
BACKGROUND: 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.
RESULTS: 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.
CONCLUSIONS: 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.
RESULTS: 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.
CONCLUSIONS: 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.
摘要：
背景：鼠疫耶尔森氏菌是一种引起鼠疫的细菌。它在整个历史上造成了许多人的死亡。该细菌具有几种毒力因子(pPla，pFra,和PYV）。PFra质粒编码部分1(F1)荚膜抗原。F1蛋白通过吞噬过程保护细菌免受宿主免疫细胞的侵害。该蛋白质对鼠疫耶尔森氏菌具有特异性。许多诊断技术基于不同食物和临床样品中F1蛋白的分子和血清学检测和定量。适体是可以作为许多靶标的特异性配体的小核酸序列。这项研究,旨在分离针对F1蛋白的高亲和力ssDNA适体。
结果：在这项研究中，SELEX被用作筛选适体的主要策略。此外，使用酶联适体吸附测定（ELASA）和表面等离子体共振（SPR）来确定获得的适体对F1蛋白的亲和力和特异性。分析表明，在获得的适体中，选择了Yer21，Yer24和Yer25的三个适体，其KD值为1.344E-7，2.004E-8和1.68E-8M，分别。对于Yer21，Yer24和Yer25，检测限（LoD）分别为0.05、0.076和0.033μg/ml。
结论：这项研究表明，合成的适体可以作为检测和分析F1蛋白的有效工具，表明它们在未来诊断应用中的潜在价值。
结果：在这项研究中，SELEX被用作筛选适体的主要策略。此外，使用酶联适体吸附测定（ELASA）和表面等离子体共振（SPR）来确定获得的适体对F1蛋白的亲和力和特异性。分析表明，在获得的适体中，选择了Yer21，Yer24和Yer25的三个适体，其KD值为1.344E-7，2.004E-8和1.68E-8M，分别。对于Yer21，Yer24和Yer25，检测限（LoD）分别为0.05、0.076和0.033μg/ml。
结论：这项研究表明，合成的适体可以作为检测和分析F1蛋白的有效工具，表明它们在未来诊断应用中的潜在价值。
