Chicken immunoglobulin Y based FRET assay for TSST-1 detection and its validation onto clinical isolates
| Autor: | S. Naveen, J. Achuth, Krishna Kadirvelu, R. M. Renuka |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Virulence
02 engineering and technology 010402 general chemistry medicine.disease_cause 01 natural sciences Microbiology Antibiotic resistance Materials Chemistry medicine Superantigen Electrical and Electronic Engineering Instrumentation biology Chemistry Toxin Metals and Alloys 021001 nanoscience & nanotechnology Condensed Matter Physics 0104 chemical sciences Surfaces Coatings and Films Electronic Optical and Magnetic Materials Förster resonance energy transfer Staphylococcus aureus biology.protein Immunoglobulin Y Antibody 0210 nano-technology |
| Zdroj: | Sensors and Actuators B: Chemical. 291:102-112 |
| ISSN: | 0925-4005 |
| DOI: | 10.1016/j.snb.2019.04.069 |
| Popis: | Staphylococcus aureus (S. aureus) is commensal bacteria implicated in various opportunistic infections worldwide. Their numerous virulence factors along with mounting antibiotic resistance is an ominous problem associated with its clinical management. Among the numerous S. aureus virulence factors, toxic shock syndrome toxin 1 (TSST-1) is a superantigen that is pivotal in life-threatening multisystem disorder. The present study reports the development of a fluorescence resonance energy transfer (FRET) assay with hybrid gold nanoparticles (GNP) conjugated chicken antibodies (IgY) and Cadmium Telluride (CdTe) quantum dots labeled rabbit antibodies for TSST-1 detection. The TSST-1 gene cloned into pET22b (+) vector for recombinant protein production and employed for anti-TSST-1 antibody generation from chicken and rabbit systems. Thereafter, CdTe-QDs tagged anti-TSST-1 IgG was employed as the fluoroprobe and subsequently, GNP conjugated anti-TSST-1 IgY served as its quencher pair. Further analysis results showed that the assay was simple, sensitive and reliable with a limit of detection (LOD) of 0.0005 μgmL-1 and shows no cross-reactivity towards closely related toxins. The artificially spiked clinical sample analysis revealed insignificant matrix interference as well as considerable toxin recovery range (81.21%–99.3%) for the assay. Furthermore, its co-evaluation with commercially available TSST-1 detection kit demonstrated their feasibility for toxin screening with substantially less assay duration. Therefore, the assay may aid in better toxic shock management, apparently reducing empirical treatment, medical costs, and antimicrobial resistance. |
| Databáze: | OpenAIRE |
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