Rapid and Sensitive Detection of Troponin I-T-C Complex from Human Serum using Microwave-Accelerated Metal-Enhanced Fluorescence

Kadir Aslan

doi:10.5101/nbe.v3i3.p179-183

Abstract

We present the rapid and sensitive detection of Troponin I-T-C (Tn I-T-C) complex from buffer and human serum samples using Microwave-Accelerated and Metal-Enhanced Fluorescence (MA-MEF) technique, which is based on the combined use of low power microwave heating, silver nanoparticle films (SNFs) and fluorescence spectroscopy. The detection of Tn I-T-C complex from buffer solutions and human serum samples on SNFs was carried out using fluorescence-based immunoassays at room temperature (control immunoassay, 2 hour total assay time) and using low-power microwave heating (MA-MEF-based immunoassay, 1 minute total assay time). A lower detection limit for Tn I-T-C complex from buffer solutions in the control immunoassay and MA-MEF-based immunoassay was 0.01 ng/ml and 0.005 ng/ml, respectively. However, the lower detection limit for Tn I-T-C complex from human serum in the control immunoassay was increased to 10 ng/ml. The use of MA-MEF technique afforded for the detection of Tn I-T-C complex from human serum samples in 1 min with a lower detection limit of 0.05 ng/ml.

Highlights

Cardiovascular diseases, such as myocardial infarction (AMI), are among the leading causes of mortality in developed countries
The detection of Tn I-T-C complex from buffer solutions and human serum samples on silver nanoparticle films (SNFs) was carried out using fluorescence-based immunoassays at room temperature (control immunoassay and using low-power microwave heating (MA-MEF-based immunoassay)
Fluorescence emission spectra reveal that both immunoassays yield similar spectra for the range of concentrations of Tn I-T-C (1 pg ml-1 - 100 ng ml-1)

Summary

Introduction

Cardiovascular diseases, such as myocardial infarction (AMI), are among the leading causes of mortality in developed countries. The stability of cTnI is significantly improved in binary complex formed with TnC or in the ternary complex cTnI-cTnT-TnC [6] In this regard, one can potentially better assess AMI by testing the human serum samples for these troponin complexes. Our research group has previously reported the rapid detection of TnI from human whole blood samples using MA-MEF technique [7] In this previous paper [7], the detection of TnI from buffer samples was first carried out using a sandwich-type immunoassay on SNFs at room temperature (control immunoassay) and using microwave heating (MA-MEF-based immunoassay), where the lower detection limit for TnI was 0.1 ng ml -1 and 0.005 ng ml-1, respectively. The use of MA-MEF technique afforded for the detection of Tn I-T-C complex from human serum samples in 1 min with a lower detection limit of 0.05 ng ml-1

Materials

Methods

Results and discusion