Abstract

Quorum sensing (QS) is a bacterial cell density-based communication system using low molecular weight signals called autoinducers (AIs). Identification and quantification of these molecules could provide valuable information related to the stage of colonization or infection as well as the stage of the disease. With this scenario, we report here for the first time the development of antibodies against the PQS (pseudomonas quinolone signal), the main signaling molecule from the pqs QS system of Pseudomonas aeruginosa, and the development of a microplate-based enzyme-linked immunosorbent assay (ELISA) able of quantifying this molecule in complex biological media in the low nanometer range (LOD, 0.36 ± 0.14 nM in culture broth media). Moreover, the PQS ELISA here reported has been found to be robust and reliable, providing accurate results in culture media. The technique allowed us to follow up the PQS profile of the release of bacterial clinical isolates obtained from patients of different disease status. A clear correlation was found between the PQS immunoreactivity equivalents and the chronic or acute infection conditions, which supports the reported differences on virulence and behavior of these bacterial strains due to their adaptation capability to the host environment. The results obtained point to the potential of the PQS as a biomarker of infection and to the value of the antibodies and the technology developed for improving diagnosis and management of P. aeruginosa infections based on the precise identification of the pathogen, appropriate stratification of the patients according to their disease status, and knowledge of the disease progression.

Highlights

  • An Immunochemical Approach to Quantify and Assess the Potential Value of Pseudomonas Quinolone Signal (PQS) as a Biomarker of Infection

  • Liquid chromatography/electrospray ionization/mass spectrometry (LC/ESI/MS) was performed in a Waters (Milford, MA, USA) model composed by an Acquity UPLC system directly interfaced to a Micromass LCT Premier XE MS system equipped with an ESI LockSpray source for monitoring positive and negative ions

  • A PQS was used to build standard curves which were measured with the different As/bioconjugate combinations, using the same procedure reported in the main body of the article for the As385/HHQ-BSA ELISA, but with the concentrations of immunoreagents shown in the table, and using PBST in the competition step

Read more

Summary

Supporting Information

An Immunochemical Approach to Quantify and Assess the Potential Value of Pseudomonas Quinolone Signal (PQS) as a Biomarker of Infection. A. Nanobiotechnology for diagnostics (Nb4D), Department of of Surfactants and Nanobiotechnology, Institute for Advanced Chemistry of Catalonia (IQAC) of the Spanish Council for Scientific Research (CSIC), Spain. B. CIBER de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN), Jordi Girona 18-26, 08034 Barcelona, Spain. C. Microbiology Department, Vall d’Hebron University Hospital (VHUH), Barcelona, Spain D. Genetics and Microbiology Department, Universitat Autònoma de Barcelona (UAB), Barcelona, Spain

GENERAL MATERIALS AND METHODS
HAPTEN DENSITY ANALYSIS OF THE BIOCONJUGATES
Hapten density
Hill Slope
Amin Amax Slope
Findings
CFUs acute
Full Text
Published version (Free)

Talk to us

Join us for a 30 min session where you can share your feedback and ask us any queries you have

Schedule a call