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High-Throughput 2017, 6(4), 14; doi:10.3390/ht6040014

Development of a Bead-Based Multiplex Assay for the Analysis of the Serological Response against the Six Pathogens HAV, HBV, HCV, CMV, T. gondii, and H. pylori

1
NMI Natural and Medical Sciences Institute at the University of Tuebingen, 72770 Reutlingen, Germany
2
TWINCORE, Centre for Experimental and Clinical Infection Research, 30625 Hanover, Germany
3
Helmholtz Centre for Infection Research, 38124 Brunswick, Germany
4
Centre for Individualized Infection Medicine, 30625 Hanover, Germany
5
Translational Infrastructure Epidemiology, German Centre for Infection Research (DZIF), 30625 Hanover, Germany
6
Immunobiology of Dendritic Cells, Institute Pasteur, 75015 Paris, France
7
Inserm U1223, Institute Pasteur, 75015 Paris, France
8
Centre for Translational Research, Institute Pasteur, 75015 Paris, France
9
Institut für Mikrobiologie und Hygiene, 66421 Homburg/Saar, Germany
10
TUM Technical University of Munich, 80333 Munich, Germany
*
Author to whom correspondence should be addressed.
Academic Editors: Xiaobo Yu and Joshua LaBaer
Received: 31 August 2017 / Revised: 17 October 2017 / Accepted: 26 October 2017 / Published: 30 October 2017
(This article belongs to the Special Issue Protein Microarrays)
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Abstract

The spread of infectious diseases and vaccination history are common subjects of epidemiological and immunological research studies. Multiplexed serological assays are useful tools for assessing both current and previous infections as well as vaccination efficacy. We developed a serological multi-pathogen assay for hepatitis A, B and C virus, cytomegalovirus (CMV), Toxoplasma gondii, and Helicobacter pylori using a bead-based multiplex assay format. The multi-pathogen assay consisting of 15 antigens was utilized for the analysis of the serological response in elderly individuals of an influenza vaccination study (n = 34). The technical assay validation revealed a mean intra-assay precision of coefficient of variation (CV) = 3.2 ± 1.5% and a mean inter-assay precision of CV = 8.2 ± 5.3% across all 15 antigens and all tested samples, indicating a robust test system. Furthermore, the assay shows high sensitivities (ranging between 94% and 100%) and specificities (ranging between 93% and 100%) for the different pathogens. The highest seroprevalence rates in our cohort were observed for hepatitis A virus (HAV; 73.5%), followed by CMV (70.6%), T. gondii (67.6%) and H. pylori (32.4%). Seroprevalences for hepatitis B virus (HBV, 8.8%) and hepatitis C virus (HCV, 0%) were low. The seroprevalences observed in our study were similar to those from other population-based studies in Germany. In summary, we conclude that our multiplex serological assay represents a suitable tool for epidemiological studies. View Full-Text
Keywords: multiplex; serotest; multi-pathogen assay; seroprevalence; hepatitis; cytomegalovirus (CMV); Toxoplasma gondii; Helicobacter pylori multiplex; serotest; multi-pathogen assay; seroprevalence; hepatitis; cytomegalovirus (CMV); Toxoplasma gondii; Helicobacter pylori
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Filomena, A.; Pessler, F.; Akmatov, M.K.; Krause, G.; Duffy, D.; Gärtner, B.; Gerhard, M.; Albert, M.L.; Joos, T.O.; Schneiderhan-Marra, N. Development of a Bead-Based Multiplex Assay for the Analysis of the Serological Response against the Six Pathogens HAV, HBV, HCV, CMV, T. gondii, and H. pylori. High-Throughput 2017, 6, 14.

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