Performance of a Multiplex Serological Helicobacter pylori Assay on a Novel Microfluidic Assay Platform

Abstract

Infection with <i>Helicobacter pylori</i> (<i>H. pylori</i>) occurs in 50% of the world population, and is associated with the development of ulcer and gastric cancer. Serological diagnostic tests indicate an <i>H. pylori</i> infection by detecting antibodies directed against <i>H. pylori</i> proteins. In addition to line blots, multiplex assay platforms provide smart solutions for the simultaneous analysis of antibody responses towards several <i>H. pylori</i> proteins. We used seven <i>H. pylori</i> proteins (FliD, gGT, GroEL, HpaA, CagA, VacA, and HP0231) and an <i>H. pylori</i> 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 <i>H. pylori</i>-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.