Immunochemical Determination of Pyocyanin and 1-Hydroxyphenazine as Potential Biomarkers of <i>Pseudomonas aeruginosa</i> Infections

Pastells, Carme; Pascual, Núria; Sánchez‐Baeza, Francisco; Marco, M.‐Pilar

doi:10.1021/acs.analchem.5b03490

Cited by 34 publications

(33 citation statements)

References 42 publications

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“…The antimicrobial activity of OBPs could also be exerted by scavenging other compounds produced by pathogenic microorganisms, with physico-chemical properties and/or molecular masses comparable to those of the ligands of these proteins. As an example, the binding tests for pyocyanin showed that it can be significantly scavenged by submillimolar levels of bovine and porcine OBPs at concentration levels higher than 5 μM, being this concentration value comparable to those active in vivo during PA infections [39, 40 and 41]. The binding capacity of OBPs against pyocyanin is an additional indication of the potential scavenging-based antimicrobial activity of the protein against PA. Further studies dealing with other bacterial and yeast toxins, as well as with diffusible bioactive compounds having molecular characteristics comparable to those of the OBP ligands, could corroborate the hypothesis that OBPs might exert an in vivo scavenging-based antimicrobial activity against a wide spectrum of pathogenic microorganisms.…”

Section: Discussionmentioning

confidence: 99%

Vertebrate odorant binding proteins as antimicrobial humoral components of innate immunity for pathogenic microorganisms

et al. 2019

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The bacterium Pseudomonas aeruginosa (PA) and the yeast Candida albicans (CA) are pathogens that cohabit the mucosa of the respiratory tracts of animals and humans. Their virulence is largely determined by chemical communication driven by quorum sensing systems (QS), and the cross perception of their quorum sensing molecules (QSM) can modulate the prevalence of one microorganism over the other. Aiming to investigate whether some of the protein components dissolved in the mucus layering the respiratory mucosa might interfere with virulence and cross-communication of these, and eventually other microorganisms, ligand binding assays were carried out to test the scavenging potential of the bovine and porcine forms of the Lipocalin odorant binding protein (OBP) for several QSMs (farnesol, and acylhomoserine lactones), and for pyocyanin, a toxin produced by PA. In addition, the direct antimicrobial activity of the OBPs was tested by time kill assay (TKA) against CA, PA and other bacteria and yeasts. The positivity of all the ligand binding assays and the antimicrobial activity determined for CA, and for some of the other microorganisms tested, let hypothesize that vertebrate OBPs might behave as humoral components of innate immunity, active against pathogenic bacteria and fungi. In addition, TKAs with mutants of bovine OBP with structural properties different from those of the native form, and with OBP forms tagged with histidines at the amino terminal, provided information about the mechanisms responsible of their antimicrobial activity and suggested possible applications of the OBPs as alternative or co-adjuvants to antibiotic therapeutic treatments.

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Section: Discussionmentioning

confidence: 99%

Vertebrate odorant binding proteins as antimicrobial humoral components of innate immunity for pathogenic microorganisms

et al. 2019

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“…PYO was obtained from Sigma-Aldrich Co. (St. Louis, MO), and 1-OHphz was synthesized following the procedures ( Vivian, 1956 ). The development of the anti-1-OHphz pAb As230 was described by Pastells et al. (2016) .…”

Section: Methodsmentioning

confidence: 99%

“…These methods, based on the specific interaction between an antigen and an antibody (Ab), are widely implemented in clinical laboratories ( Ma et al., 2019 ; Vatankhah et al., 2019 ; Zelini et al., 2019 ). Few years ago, we reported for the first time the development of polyclonal antibodies (pAbs) and microplate-based ELISA for 1-hydroxyphenazine (1-OHphz), the main metabolite of PYO, with an excellent detectability (IC 50 = 0.53 nM) ( Pastells et al., 2016 ). It has been reported that 1-OHphz also contributes to the virulence of P. aeruginosa .…”

Section: Introductionmentioning

confidence: 99%

Diagnosis and Stratification of Pseudomonas aeruginosa Infected Patients by Immunochemical Quantitative Determination of Pyocyanin From Clinical Bacterial Isolates

Rodriguez-Urretavizcaya

Pascual

Pastells

et al. 2021

Front. Cell. Infect. Microbiol.

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The development of a highly sensitive, specific, and reliable immunochemical assay to detect pyocyanin (PYO), one of the most important virulence factors (VFs) of Pseudomonas aeruginosa, is here reported. The assay uses a high-affinity monoclonal antibody (mAb; C.9.1.9.1.1.2.2.) raised against 1-hydroxyphenazine (1-OHphz) hapten derivatives (PC1; a 1:1 mixture of 9-hydroxy- and 6-hydroxy-phenazine-2-carobxylic acids). Selective screening using PYO and 1-OHphz on several cloning cycles allowed the selection of a clone able to detect PYO at low concentration levels. The microplate-based ELISA developed is able to achieve a limit of detection (LoD) of 0.07 nM, which is much lower than the concentrations reported to be found in clinical samples (130 μM in sputa and 2.8 μM in ear secretions). The ELISA has allowed the investigation of the release kinetics of PYO and 1-OHphz (the main metabolite of PYO) of clinical isolates obtained from P. aeruginosa-infected patients and cultured in Mueller–Hinton medium. Significant differences have been found between clinical isolates obtained from patients with an acute or a chronic infection (~6,000 nM vs. ~8 nM of PYO content, respectively) corroborated by the analysis of PYO/1-OHphz levels released by 37 clinical isolates obtained from infected patients at different stages. In all cases, the levels of 1-OHphz were much lower than those of PYO (at the highest levels 6,000 nM vs. 300 nM for PYO vs. 1-OHphz, respectively). The results found point to a real potential of PYO as a biomarker of P. aeruginosa infection and the possibility to use such VF also as a biomarker for patient stratification[2] and for an effective management of these kinds of infections.

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“…In a previous study, Pastells and co-workers also published an immunochemical method for the quantification of PYO and its precursor 1-hydroxyphenazine (1-OHphz) in bacterial cultures and sputum samples [133]. PYO quantification could be done in less than 2 h after conversion to 1-OHphz under basic conditions, presenting a LOD of 0.6 nM.…”

Section: Immunochemical-based Techniquesmentioning

confidence: 99%

Biological and clinical significance of quorum sensing alkylquinolones: current analytical and bioanalytical methods for their quantification

Montagut

Marco

2021

Anal Bioanal Chem

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Quorum sensing (QS) is a sophisticated bacterial communication system which plays a key role in the virulence and biofilm formation of many pathogens. The Pseudomonas aeruginosa QS network consists of four sets of connected systems (las, rlh, pqs and iqs) hierarchically organized. The pqs system involves characteristic autoinducers (AI), most of them sharing an alkylquinolone (AQ) structure, and is able to carry out several relevant biological functions besides its main signalling activity. Their role in bacterial physiology and pathogenicity has been widely studied. Indeed, the presence of these metabolites in several body fluids and infected tissues has pointed to their potential value as biomarkers of infection. In this review, we summarize the most recent findings about the biological implications and the clinical significance of the main P. aeruginosa AQs. These findings have encouraged the development of analytical and bioanalytical techniques addressed to assess the role of these metabolites in bacterial growth and survival, during pathogenesis or as biomarkers of infections. The availability of highly sensitive reliable analytical methods suitable for clinical analysis would allow getting knowledge about pathogenesis and disease prognosis or progression, supporting clinicians on the decision-making process for the management of these infections and guiding them on the application of more effective and appropriate treatments. The benefits from the implementation of the pointof-care (PoC)-type testing in infectious disease diagnostics, which are seen to improve patient outcomes by promoting earlier therapeutic interventions, are also discussed.

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Immunochemical Determination of Pyocyanin and 1-Hydroxyphenazine as Potential Biomarkers of Pseudomonas aeruginosa Infections

Cited by 34 publications

References 42 publications

Vertebrate odorant binding proteins as antimicrobial humoral components of innate immunity for pathogenic microorganisms

Vertebrate odorant binding proteins as antimicrobial humoral components of innate immunity for pathogenic microorganisms

Diagnosis and Stratification of Pseudomonas aeruginosa Infected Patients by Immunochemical Quantitative Determination of Pyocyanin From Clinical Bacterial Isolates

Biological and clinical significance of quorum sensing alkylquinolones: current analytical and bioanalytical methods for their quantification

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