Isolation and characterization of ssDNA aptamers against BipD antigen of Burkholderia pseudomallei.
| Autor: | Selvam K; Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Kubang Kerian, 16150, Kelantan, Malaysia., Najib MA; Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Kubang Kerian, 16150, Kelantan, Malaysia., Khalid MF; Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Kubang Kerian, 16150, Kelantan, Malaysia., Yunus MH; Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Kubang Kerian, 16150, Kelantan, Malaysia., Wahab HA; School of Pharmaceutical Sciences, Universiti Sains Malaysia, 11800, Pulau, Pinang, Malaysia., Harun A; Department of Medical Microbiology and Parasitology, School of Medical Sciences, Universiti Sains Malaysia, Kubang Kerian, 16150, Kelantan, Malaysia; Hospital Universiti Sains Malaysia, Kubang Kerian, 16150, Kelantan, Malaysia., Zainulabid UA; Department of Internal Medicine, Kulliyyah of Medicine, International Islamic University Malaysia, Kuantan, 25200, Pahang, Malaysia., Fadzli Mustaffa KM; Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Kubang Kerian, 16150, Kelantan, Malaysia., Aziah I; Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Kubang Kerian, 16150, Kelantan, Malaysia. Electronic address: aziahismail@usm.my. |
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| Jazyk: | angličtina |
| Zdroj: | Analytical biochemistry [Anal Biochem] 2024 Dec; Vol. 695, pp. 115655. Date of Electronic Publication: 2024 Aug 28. |
| DOI: | 10.1016/j.ab.2024.115655 |
| Abstrakt: | Background: Melioidosis is difficult to diagnose due to its wide range of clinical symptoms. The culture method is time-consuming and less sensitive, emphasizing the importance of rapid and accurate diagnostic tests for melioidosis. Burkholderia invasion protein D (BipD) of Burkholderia pseudomallei is a potential diagnostic biomarker. This study aimed to isolate and characterize single-stranded DNA aptamers that specifically target BipD. Methods: The recombinant BipD protein was produced, followed by isolation of BipD-specific aptamers using Systematic Evolution of Ligands by EXponential enrichment. The binding affinity and specificity of the selected aptamers were evaluated using Enzyme-Linked Oligonucleotide Assay. Results: The fifth SELEX cycle showed a notable enrichment of recombinant BipD protein-specific aptamers. Sequencing analysis identified two clusters with a total of seventeen distinct aptamers. AptBipD1, AptBipD13, and AptBipD50 were chosen based on their frequency. Among them, AptBipD1 exhibited the highest binding affinity with a K Conclusion: AptBipD1 is a promising candidate for further development of reliable, affordable, and efficient point-of-care diagnostic tests for melioidosis. Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. (Copyright © 2024 Elsevier Inc. All rights reserved.) |
| Databáze: | MEDLINE |
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