title: Selection of DNA aptamers against Human Cardiac Troponin I for colorimetric sensor based dot blot application.
abstract: Troponin T and I are ideal markers which are highly sensitive and specific for myocardial injury and have shown better efficacy than earlier markers. Since aptamers are ssDNA or RNA that bind to a wide variety of target molecules, the purpose of this research was to select an aptamer from a 79bp single-stranded DNA (ssDNA) random library that was used to bind the Human Cardiac Troponin I from a synthetic nucleic acids library by systematic evolution of ligands exponential enrichment (Selex) based on several selection and amplification steps. Human Cardiac Troponin I protein was coated onto the surface of streptavidin magnetic beads to extract specific aptamer from a large and diverse random ssDNA initial oligonucleotide library. As a result, several aptamers were selected and further examined for binding affinity and specificity. Finally TnIApt 23 showed beast affinity in nanomolar range (2.69nM) toward the target protein. A simple and rapid colorimetric detection assay for Human Cardiac Troponin I using the novel and specific aptamer-AuNPs conjugates based on dot blot assay was developed. The detection limit for this protein using aptamer-AuNPs-based assay was found to be 5ng/ml.
authors.affiliation: Applied Biotechnology Research Center, Baqiyatallah University of Medical Science, Tehran, Iran. Electronic address: Dorraj.gh@gmail.com.
authors.forename: Ghamar Soltan
authors.initials: GS
authors.lastname: Dorraj
authors.affiliation: Medical Biotechnology Department, Tarbiat Modares University, Tehran, Iran. Electronic address: rasaee_mj@yahoo.com.
authors.forename: Mohammad Javad
authors.initials: MJ
authors.lastname: Rassaee
authors.affiliation: Applied Biotechnology Research Center, Baqiyatallah University of Medical Science, Tehran, Iran. Electronic address: amlatifi290@gmail.com.
authors.forename: Ali Mohammad
authors.initials: AM
authors.lastname: Latifi
authors.affiliation: Department of Cardiology and Cardiothoracic Surgery, Baqiyatallah University of Medical Sciences, Tehran, Iran. Electronic address: Pishgoobm@yahoo.com.
authors.forename: Bahram
authors.initials: B
authors.lastname: Pishgoo
authors.affiliation: Human Genetic Research Center, Baqiyatallah University of Medical Science, Tehran, Iran. Electronic address: Tavalla.mah@gmail.com.
authors.forename: Mahmood
authors.initials: M
authors.lastname: Tavallaei
mesh_terms: D052157:Aptamers, Nucleotide / Q000737:chemistry*; D015374:Biosensing Techniques / Q000379:methods*; D003124:Colorimetry / Q000379:methods; D006046:Gold / Q000737:chemistry*; D006801:Humans; D053768:Metal Nanoparticles / Q000737:chemistry*; D052156:SELEX Aptamer Technique / Q000379:methods; D019210:Troponin I / Q000032:analysis*
chemical_list: D052157:Aptamers, Nucleotide; D019210:Troponin I; D006046:Gold
keywords: DNA aptamer; Dot blot assay; Gold nanoparticle; Human Cardiac Troponin I; Selex