Development of a Point-of-Care System Based on White Light Reflectance Spectroscopy: Application in CRP Determination.

Dimitra Tsounidi,Evangelia Legaki,Konstantina Panagiotopoulou,Styliani Taka,Sotirios Kakabakos,Panagiota Petrou,Vasiliki Giogli,Ioannis Raptis,Georgios Koukouvinos,Zoi Ekaterinidi,Vasilios Christianidis

doi:10.3390/bios11080268

Dimitra Tsounidi, Evangelia Legaki + Show 9 more

Open Access

https://doi.org/10.3390/bios11080268

Copy DOI

Abstract

The development of methods and miniaturized systems for fast and reliable quantitative determinations at the Point-of-Care is a top challenge and priority in diagnostics. In this work, a compact bench-top system, based on White Light Reflectance Spectroscopy, is introduced and evaluated in an application with high clinical interest, namely the determination of C-Reactive protein (CRP) in human blood samples. The system encompassed all the necessary electronic and optical components for the performance of the assay, while the dedicated software provided the sequence and duration of assay steps, the reagents flow rate, the real-time monitoring of sensor response, and data processing to deliver in short time and accurately the CPR concentration in the sample. The CRP assay included two steps, the first comprising the binding of sample CRP onto the chip immobilized capture antibody and the second the reaction of the surface immunosorbed CRP molecules with the detection antibody. The assay duration was 12 min and the dynamic range was from 0.05 to 200 μg/mL, covering both normal values and acute inflammation incidents. There was an excellent agreement between CRP values determined in human plasma samples using the developed device with those received for the same samples by a standard diagnostic laboratory method.

Highlights

C-Reactive Protein (CRP), a 118 kDa molecular weight protein, is the most established biomarker of inflammation
C-Reactive Protein (CRP) from human fluids were purchased from Scripps Laboratories (San Diego, CA, USA)
The detection of CRP in human plasma using the proposed White Light Reflectance Spectroscopy (WLRS) device is based on a label-free two-site sandwich immunoassay

Summary

Introduction

C-Reactive Protein (CRP), a 118 kDa molecular weight protein, is the most established biomarker of inflammation. It consists of five non-covalently bound identical subunits and belongs to the class of acute phase proteins [1]. CRP can be used to distinguish bacterial from viral infection since its levels are much higher in case of bacterial infections, rather than in viral ones. It is widely used as a biomarker for sepsis diagnosis, which is one of the major mortality causes in critically ill patients [4,5]

Methods

Results

Conclusion