Chicken immunoglobulin Y based FRET assay for TSST-1 detection and its validation onto clinical isolates

Publication date: 15 July 2019Source: Sensors and Actuators B: Chemical, Volume 291Author(s): J. Achuth, R.M. Renuka, S. Naveen, K. KadirveluAbstractStaphylococcus 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 as...
Source: Sensors and Actuators B: Chemical - Category: Chemistry Source Type: research