Molecularly imprinted polymers as effective capturing receptors in a pseudo-ELISA immunoassay for procalcitonin detection in veterinary species.

The research article introduces a new immunoassay method based on molecularly imprinted polymers that can accurately measure procalcitonin, a biomarker of sepsis, in animal species.

Research Overview

The research focuses on the development of a new immunoenzymatic assay, or a biochemical test, that uses a molecularly imprinted polymer (MIP) as an artificial antibody. This test, known as a pseudo-ELISA, was designed to measure the quantity of procalcitonin (PCT), a calcium-regulating hormone, in animal species.

• The measurement of PCT in human medical practices is a critical diagnostic tool for sepsis, a life-threatening condition that arises when an infection triggers a violent immune response in the body. However, there are very few clinical studies that examine the relevance of PCT as an indicator of sepsis in veterinary patients, largely due to the lack of validated testing methods.
• MIPs, a system where specific particles can be created to match those of the targeted molecule, have been useful substitutes for actual antibodies in several important applications. As such, MIP-based sandwich assays have become promising analytical materials for the detection of disease biomarkers, substances or processes in the body that can indicate the presence of disease.

Methodology and Results

In this experiment, the researchers synthesized a polynorepinephrine (PNE)-based imprinted film directly onto the surface of a well in a 96-well plate. With this setup, the team then measured the quantity of PCT in the tested samples through a colorimetric sandwich assay, a technique used to detect the presence of a substance in a solution, by replacing the kit’s capture antibody with the PNE-based MIP.

• The pseudo-ELISA was used to detect PCT in both canine and equine specimens in buffer and plasma solutions.
• When carried out under optimized conditions, the test results from the plasma samples showed a limit of detection (LOD) of 5.87 ng/mL and a coefficient of variation (CV), or reproducibility rate, of 10% for the canine samples.
• For the equine samples, a LOD of 4.46 ng/mL and a CV of 7.61% were recorded.

This study emphasizes the potential of using MIP-based immunoassays for reliable and efficient detection of sepsis biomarkers in animal species. This development could pave the way for better diagnostics and treatment plans for sepsis in veterinary medicine.