Isolation and characterization of ssDNA aptamers against BipD antigen of Burkholderia pseudomallei

Selvam, Kasturi and Najib, Mohamad Ahmad and Khalid, Muhammad Fazli and Yunus, Muhammad Hafiznur and A Wahab, Habibah and Harun, Azian and Zainulabid, Ummu Afeera and Mustaffa, Khairul Mohd Fadzli and Ismail, Aziah (2024) Isolation and characterization of ssDNA aptamers against BipD antigen of Burkholderia pseudomallei. Analytical Biochemistry, 695. pp. 1-11. ISSN 0003-2697 E-ISSN 1096-0309

	PDF (Journal) - Published Version Restricted to Repository staff only Download (4MB) | Request a copy
Preview	PDF (Scopus) - Supplemental Material Download (268kB) | Preview
Preview	PDF (WOS) - Supplemental Material Download (331kB) | Preview

Abstract

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 d value of 1.0 μ M for the recombinant BipD protein. Furthermore, AptBipD1 showed significant specificity for B. pseudomallei compared to other tested bacteria. Conclusion: AptBipD1 is a promising candidate for further development of reliable, affordable, and efficient point-of-care diagnostic tests for melioidosis.

Item Type:	Article (Journal)
Additional Information:	7459/115497
Uncontrolled Keywords:	Melioidosis; B. pseudomallei; BipD; ssDNA ; aptamers; SELEX
Subjects:	R Medicine > RC Internal medicine > RC111 Infectious and Parasitic Diseases
Kulliyyahs/Centres/Divisions/Institutes (Can select more than one option. Press CONTROL button):	Kulliyyah of Medicine > Department of Internal Medicine Kulliyyah of Medicine
Depositing User:	Asst. Dr. Ummu Afeera Zainulabid
Date Deposited:	05 Nov 2024 14:25
Last Modified:	12 Dec 2024 10:34
URI:	http://irep.iium.edu.my/id/eprint/115497

Actions (login required)

View Item