Abstract
To achieve improved sensitivity in cardiac biomarker detection, a batch incubation magnetic microbead immunoassay was developed and tested on three separate human protein targets: myoglobin, heart-type fatty acid binding protein, and cardiac troponin I. A sandwich immunoassay was performed in a simple micro-centrifuge tube allowing full dispersal of the solid capture surface during incubations. Following magnetic bead capture and wash steps, samples were analyzed in the presence of a manipulated magnetic field utilizing a modified microscope slide and fluorescent inverted microscope to collect video data files. Analysis of the video data allowed for the quantitation of myoglobin, heart-type fatty acid binding protein and cardiac troponin I to levels of 360 aM, 67 fM, and 42 fM, respectively. Compared to the previous detection limit of 50 pM for myoglobin, this offers a five-fold improvement in sensitivity. This improvement in sensitivity and incorporation of additional markers, along with the small sample volumes required, suggest the potential of this platform for incorporation as a detection method in a total sample analysis device enabling multiplexed detection for the analysis of clinical samples.
Highlights
Due to their high sensitivity, biosensors have become a popular diagnostic tool for both early and rapid disease detection
The American College of Cardiology (ACC) and the American Heart Association (AHA) currently recognize a biomarker panel composed of myoglobin, cardiac troponin I, and creatine kinase MB (CK-MB) for the diagnosis of acute myocardial infarction (AMI).[2,3]
Because CK-MB has a low sensitivity for AMI within six hours a er an incident and cardiac troponin I (cTnI) is better at detecting minor cardiac damage, CK-MB was not evaluated in this study.[2]
Summary
Sensitive detection of cardiac biomarkers using a magnetic microbead immunoassay Christine F. To achieve improved sensitivity in cardiac biomarker detection, a batch incubation magnetic microbead immunoassay was developed and tested on three separate human protein targets: myoglobin, hearttype fatty acid binding protein, and cardiac troponin I. Analysis of the video data allowed for the quantitation of myoglobin, heart-type fatty acid binding protein and cardiac troponin I to levels of 360 aM, 67 fM, and 42 fM, respectively. Compared to the previous detection limit of 50 pM for myoglobin, this offers a five-fold improvement in sensitivity. This improvement in sensitivity and incorporation of additional markers, along with the small sample volumes required, suggest the potential of this platform for incorporation as a detection method in a total sample analysis device enabling multiplexed detection for the analysis of clinical samples
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