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

• Main Authors: Selvam, Kasturi, Najib, Mohamad Ahmad, Khalid, Muhammad Fazli, Yunus, Muhammad Hafiznur, A Wahab, Habibah, Harun, Azian, Zainulabid, Ummu Afeera, Mustaffa, Khairul Mohd Fadzli, Ismail, Aziah
• Format: Article
• Language: English; English; English
• Published: Elsevier 2024
• Subjects: RC111 Infectious and Parasitic Diseases
• Online Access: http://irep.iium.edu.my/115497/7/115497_Isolation%20and%20characterization%20of%20ssDNA%20aptamers%20against%20BipD.pdf; http://irep.iium.edu.my/115497/13/115497_Isolation%20and%20characterization%20of%20ssDNA%20aptamers%20against%20BipD_SCOPUS.pdf; http://irep.iium.edu.my/115497/14/115497_Isolation%20and%20characterization%20of%20ssDNA%20aptamers%20against%20BipD_WOS.pdf; http://irep.iium.edu.my/115497/; https://www.sciencedirect.com/science/article/abs/pii/S0003269724001994?via%3Dihub
• Institution: Universiti Islam Antarabangsa Malaysia

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.