Innovative Aptamer-HRP Conjugation for Cardiac Troponin I Detection: A Novel ELASA Approach for AMI Diagnostics

Area of Science:

• Biochemistry
• Biotechnology
• Medical Diagnostics

Background:

• Acute myocardial infarction (AMI) is a leading cause of death globally.
• Current cardiac troponin I (cTnI) detection methods using antibodies have limitations.
• Aptamers offer a potential alternative for sensitive and specific biomarker detection.

Purpose of the Study:

• To develop and validate an aptamer-based enzyme-linked aptamer assay (ELASA) for detecting cardiac troponin I (cTnI).
• To evaluate the sensitivity, specificity, and clinical applicability of the developed aptamer-ELASA for AMI diagnosis.

Main Methods:

• Aptamer selection and characterization (Tro4 and Tro6) for cTnI binding.
• Molecular docking to predict aptamer-cTnI binding affinity.
• Development of direct and sandwich ELASA formats using biotinylated and HRP-conjugated aptamers.
• Assay validation using clinical serum samples from AMI and non-AMI patients.

Main Results:

• Molecular docking confirmed high binding affinity of Tro4 and Tro6 aptamers to cTnI.
• The sandwich ELASA demonstrated improved sensitivity and specificity for cTnI detection.
• The assay showed a linear response from 0.1-22 ng/mL cTnI with an LOD of 0.10 ng/mL.
• Results from patient samples correlated well with a commercial antibody-based assay.

Conclusions:

• A highly sensitive and specific aptamer-based sandwich ELASA for cTnI detection was successfully developed.
• The assay shows significant potential for clinical application in diagnosing AMI.
• This aptamer-ELASA presents a promising alternative to conventional antibody-based diagnostic methods.