Hydrogel based protein biochip for parallel detection of biomarkers for diagnosis of a Systemic Inflammatory Response Syndrome (SIRS) in human serum


Autoři: Anne Stumpf aff001;  Thomas Brandstetter aff001;  Johannes Hübner aff002;  Jürgen Rühe aff001
Působiště autorů: Laboratory for Chemistry and Physics of Interfaces, Department of Microsystems Engineering, University of Freiburg, Georges-Koehler-Allee, Freiburg, Germany aff001;  Division of Pediatric Infectious Diseases, Dr. von Hauner Children's Hospital, Ludwig Maximilian's University, Munich, Germany, Lindwurmstr, Munich, Germany aff002
Vyšlo v časopise: PLoS ONE 14(12)
Kategorie: Research Article
doi: 10.1371/journal.pone.0225525

Souhrn

The Systemic Inflammatory Response Syndrome (SIRS), a sepsis related inflammatory state, is a self-defense mechanism against specific and nonspecific stimuli. The six most extensively studied inflammatory biomarkers for the clinical diagnosis of SIRS are interleukin 4 (hIL-4), interleukin 6 (hIL-6), interleukin 10 (hIL-10), tumor necrosis factor alpha (hTNF-α), interferon gamma (hIFN-γ) and procalcitonin (hPCT). These biomarkers are naturally present (but usually only at low concentration) in SIRS infected patients [1, 2] and thus the development of a highly sensitive detection method is of major clinical interest. However, the existing analytical techniques are lacking in required analytical sensitivity and parallel determination of these biomarkers. We developed a fast, easy and cost-efficient protein microarray biochip where the capture molecules are attached on hydrogel spots, enabling SIRS diagnosis by parallel detection of these six clinically relevant biomarkers with a sample volume of 25 μl. With our hydrogel based protein microarray biochip we achieved a limit of detection for hIL-4 of 75.2 pg/ml, for hIL-6 of 45.1 pg/ml, for hIL-10 of 71.5 pg/ml, for hTNF-α of 56.7 pg/ml, for IFN-γ of 46.4 pg/ml and for hPCT of 1.1 ng/ml in spiked human serum demonstrating sufficient sensitivity for clinical usage. Additionally, we demonstrated successful detection of two relevant SIRS biomarkers in clinical patient samples with a turnaround time of the complete analysis from sample-to-answer in less than 200 minutes.

Klíčová slova:

Biomarkers – Enzyme-linked immunoassays – Gels – Immunoassays – Microarrays – Polymers – Serum proteins – Systemic inflammatory response syndrome


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Článek vyšel v časopise

PLOS One


2019 Číslo 12