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

Autor: Ahmad Najib M; Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Kubang Kerian, Kelantan, 16150, Malaysia., Winter A; School of Life Sciences, Keele University, Staffordshire, UK., Mustaffa KMF; Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Kubang Kerian, Kelantan, 16150, Malaysia., Ong EBB; Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Gelugor, Pulau Pinang, 11800, Malaysia., Selvam K; Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Kubang Kerian, Kelantan, 16150, Malaysia., Khalid MF; Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Kubang Kerian, Kelantan, 16150, Malaysia., Awang MS; Collaborative Microelectronic Design Excellence Centre (CEDEC), Universiti Sains Malaysia, Bayan Lepas, Pulau Pinang, 11900, Malaysia., Abd Manaf A; Collaborative Microelectronic Design Excellence Centre (CEDEC), Universiti Sains Malaysia, Bayan Lepas, Pulau Pinang, 11900, Malaysia., Bustami Y; School of Biological Sciences, Universiti Sains Malaysia, Gelugor, Pulau Pinang, 11800, Malaysia., Aziah I; Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Kubang Kerian, Kelantan, 16150, Malaysia. aziahismail@usm.my.
Jazyk: angličtina
Zdroj: Scientific reports [Sci Rep] 2024 Nov 18; Vol. 14 (1), pp. 28416. Date of Electronic Publication: 2024 Nov 18.
DOI: 10.1038/s41598-024-78685-9
Abstrakt: 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.
Competing Interests: Competing interests The authors declare no competing interests.
(© 2024. The Author(s).)
Databáze: MEDLINE
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