Isolation and characterization of DNA aptamers against the HlyE antigen of Salmonella Typhi

Ahmad Najib, Mohamad; Winter, Anja; Mustaffa, Khairul Mohd Fadzli; Ong, Eugene Boon Beng; Selvam, Kasturi; Khalid, Muhammad Fazli; Awang, Mohd Syafiq; Abd Manaf, Asrulnizam; Bustami, Yazmin; Aziah, Ismail

doi:10.1038/s41598-024-78685-9

Isolation and characterization of DNA aptamers against the HlyE antigen of Salmonella Typhi

Ahmad Najib, Mohamad; Winter, Anja; Mustaffa, Khairul Mohd Fadzli; Ong, Eugene Boon Beng; Selvam, Kasturi; Khalid, Muhammad Fazli; Awang, Mohd Syafiq; Abd Manaf, Asrulnizam; Bustami, Yazmin; Aziah, Ismail

Authors

Mohamad Ahmad Najib

Anja Winter a.winter@keele.ac.uk

Khairul Mohd Fadzli Mustaffa

Eugene Boon Beng Ong

Kasturi Selvam

Muhammad Fazli Khalid

Mohd Syafiq Awang

Asrulnizam Abd Manaf

Yazmin Bustami

Ismail Aziah

Abstract

Aptamers have emerged as prominent ligands in clinical diagnostics because they provide various advantages over antibodies, such as quicker generation time, reduced manufacturing costs, minimal batch-to-batch variability, greater modifiability, and improved thermal stability. In the present study, we isolated and characterized DNA aptamers that can specifically bind to the hemolysin E (HlyE) antigen of Salmonella Typhi for future development of typhoid diagnostic tests. The DNA aptamers against Salmonella Typhi HlyE were isolated using systematic evolution of ligands by exponential enrichment (SELEX), and their binding affinity and specificity were assessed utilizing enzyme-linked oligonucleotide assay (ELONA). A total of 11 distinct aptamers were identified, and the binding affinities and species selectivities of the three most probable aptamers were determined. Kd values were obtained in the nanomolar range, with the highest affinity of 83.6 nM determined for AptHlyE97. In addition, AptHlyE11, AptHlyE45 and AptHlyE97 clearly distinguished S. Typhi HlyE from other tested bacteria, such as Salmonella Paratyphi A, Salmonella Paratyphi B, Shigella flexneri, Klebsiella pneumonia and Escherichia coli, therefore displaying desirable specificity. These novel aptamers could be used as diagnostic ligands for the future development of inexpensive and effective point-of-care tests for typhoid surveillance, especially in developing countries of the tropics and subtropics.

Citation

Ahmad Najib, M., Winter, A., Mustaffa, K. M. F., Ong, E. B. B., Selvam, K., Khalid, M. F., …Aziah, I. (in press). Isolation and characterization of DNA aptamers against the HlyE antigen of Salmonella Typhi. Scientific Reports, 14(1), Article 28416. https://doi.org/10.1038/s41598-024-78685-9

Journal Article Type	Article
Acceptance Date	Nov 4, 2024
Online Publication Date	Nov 18, 2024
Deposit Date	Nov 26, 2024
Publicly Available Date	Nov 26, 2024
Journal	Scientific Reports
Publisher	Nature Publishing Group
Peer Reviewed	Peer Reviewed
Volume	14
Issue	1
Article Number	28416
DOI	https://doi.org/10.1038/s41598-024-78685-9
Keywords	Salmonella Typhi, SELEX, DNA aptamers, HlyE antigen
Public URL	https://keele-repository.worktribe.com/output/980700
Publisher URL	https://www.nature.com/articles/s41598-024-78685-9

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Isolation and characterization of DNA aptamers against the HlyE antigen of Salmonella Typhi (8.2 Mb)
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