Isolation and characterization of ssDNA aptamers against BipD antigen of Burkholderia pseudomallei.
Anal Biochem
; 695: 115655, 2024 Dec.
Article
em En
| MEDLINE
| ID: mdl-39214325
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.Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Proteínas de Bactérias
/
DNA de Cadeia Simples
/
Burkholderia pseudomallei
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Aptâmeros de Nucleotídeos
/
Técnica de Seleção de Aptâmeros
Limite:
Humans
Idioma:
En
Revista:
Anal Biochem
Ano de publicação:
2024
Tipo de documento:
Article
País de afiliação:
Malásia