Affiliations 

  • 1 Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Kubang Kerian, 16150, Kelantan, Malaysia
  • 2 School of Pharmaceutical Sciences, Universiti Sains Malaysia, 11800, Pulau, Pinang, Malaysia
  • 3 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
  • 4 Department of Internal Medicine, Kulliyyah of Medicine, International Islamic University Malaysia, Kuantan, 25200, Pahang, Malaysia
  • 5 Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Kubang Kerian, 16150, Kelantan, Malaysia. Electronic address: aziahismail@usm.my
Anal Biochem, 2024 Aug 28;695:115655.
PMID: 39214325 DOI: 10.1016/j.ab.2024.115655

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 Kd value of 1.0 μM 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.

* Title and MeSH Headings from MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.