Abstract
Infection with Helicobacter pylori (H. pylori) occurs in 50% of the world population, and is associated with the development of ulcer and gastric cancer. Serological diagnostic tests indicate an H. pylori infection by detecting antibodies directed against H. pylori proteins. In addition to line blots, multiplex assay platforms provide smart solutions for the simultaneous analysis of antibody responses towards several H. pylori proteins. We used seven H. pylori proteins (FliD, gGT, GroEL, HpaA, CagA, VacA, and HP0231) and an H. pylori lysate for the development of a multiplex serological assay on a novel microfluidic platform. The reaction limited binding regime in the microfluidic channels allows for a short incubation time of 35 min. The developed assay showed very high sensitivity (99%) and specificity (100%). Besides sensitivity and specificity, the technical validation (intra-assay CV = 3.7 ± 1.2% and inter-assay CV = 5.5 ± 1.2%) demonstrates that our assay is also a robust tool for the analysis of the H. pylori-specific antibody response. The integration of the virulence factors CagA and VacA allow for the assessment of the risk for gastric cancer development. The short assay time and the performance of the platform shows the potential for implementation of such assays in a clinical setting.
Highlights
EvalutionTM (MyCartis, Zwijnaarde, Belgium) is a novel microfluidic assay platform, which allows the multiplexing of up to 150 individual particle populations, combined with advantageous conditions given by the microfluidic format and a short assay time [1]
The incubation times for the samples and the detection antibody were investigated during assay
The incubation times for the samples and the detection antibody were investigated during assay development
Summary
EvalutionTM (MyCartis, Zwijnaarde, Belgium) is a novel microfluidic assay platform, which allows the multiplexing of up to 150 individual particle populations, combined with advantageous conditions given by the microfluidic format and a short assay time [1]. Serologic assays which enable the analysis of multiple antibody responses in parallel from many patients may serve as a valuable tool for population-based screening and risk assessment. Is an adhesion protein involved in colonization [13], and the H. pylori oxidoreductase DsbK regulates several bacterial virulence factors [14]. Some of these proteins had previously been employed in serologic assays [9]. Analysis of the serological response of H. pylori positive patients to individual antigens was performed with regard to their disease state
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