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| Pàgina inicial > Articles > Articles publicats > An Immunochemical Approach to Detect the Quorum Sensing-Regulated Virulence Factor 2-Heptyl-4-Quinoline N-Oxide (HQNO) Produced by Clinical Isolates |
(Centro de Investigación Biomédica en Red de Bioingeniería, Biomateriales y Nanomedicina) (Universitat Autònoma de Barcelona. Departament de Genètica i de Microbiologia) (Centro de Investigación Biomédica en Red de Bioingeniería, Biomateriales y Nanomedicina) (Centro de Investigación Biomédica en Red de Bioingeniería, Biomateriales y Nanomedicina)| Data: | 2022 |
| Resum: | Understanding quorum sensing (QS) and its role in the development of pathogenesis may provide new avenues for diagnosing, surveillance, and treatment of infectious diseases. For this purpose, the availability of reliable and efficient analytical diagnostic tools suitable to specifically detect and quantify these essential QS small molecules and QS regulated virulence factors is crucial. Here, we reported the development and evaluation of antibodies and an enzyme-linked immunosorbent assay (ELISA) for HQNO (2-heptyl-4-quinoline N-oxide), a QS product of the PqsR system, which has been found to act as a major virulence factor that interferes with the growth of other microorganisms. Despite the nonimmunogenic character of HQNO, the antibodies produced showed high avidity and the microplate-based ELISA developed could detect HQNO in the low nM range. Hence, a limit of detection (LOD) of 0. 60 ± 0. 13 nM had been reached in Müeller Hinton (MH) broth, which was below previously reported levels using sophisticated equipment based on liquid chromatography coupled to mass spectrometry. The HQNO profile of release of different clinical isolates analyzed using this ELISA showed significant differences depending on whether the clinical isolates belonged to patients with acute or chronic infections. These data point to the possibility of using HQNO as a specific biomarker to diagnose infections and for patient surveillance. Considering the role of HQNO in inhibiting the growth of coinfecting bacteria, the present ELISA will allow the investigation of these complex bacterial interactions underlying infections. IMPORTANCE Bacteria use quorum sensing (QS) as a communication mechanism that releases small signaling molecules which allow synchronizing a series of activities involved in the pathogenesis, such as the biosynthesis of virulence factors or the regulation of growth of other bacterial species. HQNO is a metabolite of the -specific QS signaling molecule PQS ( quinolone signal). In this work, the development of highly specific antibodies and an immunochemical diagnostic technology (ELISA) for the detection and quantification of HQNO was reported. The ELISA allowed profiling of the release of HQNO by clinical bacterial isolates, showing its potential value for diagnosing and surveillance of infections. Moreover, the antibodies and the ELISA reported here may contribute to the knowledge of other underlying conditions related to the pathology, such as the role of the interactions with other bacteria of a particular microbiota environment. |
| Ajuts: | Ministerio de Ciencia e Innovación SAF2015-67476-R Ministerio de Ciencia e Innovación RTI2018-096278-B-C21 Ministerio de Ciencia e Innovación BES-2016-076496 Fundació la Marató de TV3 TV32018-201825-30-31 Ministerio de Ciencia e Innovación PRE2019-087542 Agència de Gestió d'Ajuts Universitaris i de Recerca 2017/SGR-1441 |
| Drets: | Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.  |
| Llengua: | Anglès |
| Document: | Article ; recerca ; Versió publicada |
| Matèria: | Pseudomonas aeruginosa ; Quorum sensing ; Virulence ; Diagnostic ; Antibodies ; Immunoassay ; HQNO-2-heptyl-4-quinoline N-oxide ; MvfR ; PqsR ; Virulence factors |
| Publicat a: | Microbiology Spectrum, Vol. 10, Num. 4 (July 2022) , art. e01073-21, ISSN 2165-0497 |
