Design and testing of a disposable microfluidic chemiluminescent immunoassay for disease biomarkers in human serum samples

Abstract

This paper presents the development of a plastic microfluidic immunosensor for rapid, reliable and on-the-spot detection of disease biomarkers in human sera. The microfluidic chips were fabricated in cyclic polyolefin by hot-embossing with a silicon master mold. The master itself was made using photolithographic techniques and Deep Reactive Ion Etching (DRIE). As a platform model, serum concentrations of C-reactive protein (CRP), a cardiac and inflammation marker, was measured on-chip using chemiluminescence based immunoassay. The assay results were read via an on-board instant photographic film, and with an imager capable of detecting chemiluminescent signals. The on-board detection module obviates the need for any dedicated bench-top analyzer for reading the immunoassay results, and therefore makes the device self-sufficient for point-of-care diagnostics when simple positive/negative results are sought. The microfluidic chemiluminescence results were compared with standard microplate ELISA analysis to assess the accuracy of the developed microfluidic immunoassay. Screening of CRP in human serum samples showed good correlation with ELISA analysis and the mean difference between the two methods using the Bland and Altman method was -0.079 +/- 0.858 mg/L for hsCRP. With approximate assay times of 25 min, the developed microfluidic immunoassay approach shows great potential for rapid plus sensitive detection of disease markers at the point-of-care.

Biomarkers and diagnostic tools for detection of Helicobacter pylori

“…Microfluidic systems for clinical assays could also enable on-chip detection of blood-associated infectious diseases in simple, mass-producible, and thus, cheap setting (Chung et al 2009). Diagnosis of H. pylori infection could be enhanced by modification of current detection methods related to nano/microfluidics interface using lateral flow and diffusion (Bhattacharyya and Klapperich 2007;Eteshola and Leckband 2001;Morozov et al 2007). …”

“…Thus, these processes are centralized and samples should be collected and transferred to a central lab to be processed (Bhattacharyya and Klapperich 2007;Eteshola and Leckband 2001). To address these limitations, miniaturized diagnostic tools including microfluidic-based systems have been developed.…”

Microfluidic Electrochemical Detection Techniques of Cancer Biomarkers

“…Microfluidic chip is a technology platform aimed at replacing the various functions of conventional chemical or biological laboratories, which involved reaction, separation, detection and cell culture, sorting, cracking and other basic operating unit integrated or basic integrated into a few square centimeters on the chip in chemistry and biology, and other fields [23,24] ( Fig. 1).…”

“…The detection signal intensity is proportional to the concentration of the antigen. Diffusion process has a great impact on the strength of the detection signal [24][25][26][27].…”

Trends and Perspectives in Immunosensors

“…Fluorescence optical systems are preferred over light absorbance ones due to their higher sensitivity on small platforms [156]. There are also reports of small immunosensors using SPR [157,158], chemiluminescence [159], light scattering [160] and electrochemical systems [161].…”