Isolation and characterization of DNA aptamers against the HlyE antigen of Salmonella Typhi
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...
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Nature Portfolio
2024-11-01
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| Online Access: | https://doi.org/10.1038/s41598-024-78685-9 |
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| author | Mohamad Ahmad Najib Anja Winter Khairul Mohd Fadzli Mustaffa Eugene Boon Beng Ong Kasturi Selvam Muhammad Fazli Khalid Mohd Syafiq Awang Asrulnizam Abd Manaf Yazmin Bustami Ismail Aziah |
| author_facet | Mohamad Ahmad Najib Anja Winter Khairul Mohd Fadzli Mustaffa Eugene Boon Beng Ong Kasturi Selvam Muhammad Fazli Khalid Mohd Syafiq Awang Asrulnizam Abd Manaf Yazmin Bustami Ismail Aziah |
| author_sort | Mohamad Ahmad Najib |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-e5db28df31cc49528b76e897b9628b56 |
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| issn | 2045-2322 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | Nature Portfolio |
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| series | Scientific Reports |
| spelling | doaj-art-e5db28df31cc49528b76e897b9628b562025-08-20T02:22:21ZengNature PortfolioScientific Reports2045-23222024-11-0114111310.1038/s41598-024-78685-9Isolation and characterization of DNA aptamers against the HlyE antigen of Salmonella TyphiMohamad Ahmad Najib0Anja Winter1Khairul Mohd Fadzli Mustaffa2Eugene Boon Beng Ong3Kasturi Selvam4Muhammad Fazli Khalid5Mohd Syafiq Awang6Asrulnizam Abd Manaf7Yazmin Bustami8Ismail Aziah9Institute for Research in Molecular Medicine (INFORMM), Universiti Sains MalaysiaSchool of Life Sciences, Keele UniversityInstitute for Research in Molecular Medicine (INFORMM), Universiti Sains MalaysiaInstitute for Research in Molecular Medicine (INFORMM), Universiti Sains MalaysiaInstitute for Research in Molecular Medicine (INFORMM), Universiti Sains MalaysiaInstitute for Research in Molecular Medicine (INFORMM), Universiti Sains MalaysiaCollaborative Microelectronic Design Excellence Centre (CEDEC), Universiti Sains MalaysiaCollaborative Microelectronic Design Excellence Centre (CEDEC), Universiti Sains MalaysiaSchool of Biological Sciences, Universiti Sains MalaysiaInstitute for Research in Molecular Medicine (INFORMM), Universiti Sains MalaysiaAbstract 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.https://doi.org/10.1038/s41598-024-78685-9DNA aptamersSELEXHlyE antigenSalmonella Typhi |
| spellingShingle | Mohamad Ahmad Najib Anja Winter Khairul Mohd Fadzli Mustaffa Eugene Boon Beng Ong Kasturi Selvam Muhammad Fazli Khalid Mohd Syafiq Awang Asrulnizam Abd Manaf Yazmin Bustami Ismail Aziah Isolation and characterization of DNA aptamers against the HlyE antigen of Salmonella Typhi Scientific Reports DNA aptamers SELEX HlyE antigen Salmonella Typhi |
| title | Isolation and characterization of DNA aptamers against the HlyE antigen of Salmonella Typhi |
| title_full | Isolation and characterization of DNA aptamers against the HlyE antigen of Salmonella Typhi |
| title_fullStr | Isolation and characterization of DNA aptamers against the HlyE antigen of Salmonella Typhi |
| title_full_unstemmed | Isolation and characterization of DNA aptamers against the HlyE antigen of Salmonella Typhi |
| title_short | Isolation and characterization of DNA aptamers against the HlyE antigen of Salmonella Typhi |
| title_sort | isolation and characterization of dna aptamers against the hlye antigen of salmonella typhi |
| topic | DNA aptamers SELEX HlyE antigen Salmonella Typhi |
| url | https://doi.org/10.1038/s41598-024-78685-9 |
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