Abstract
Stapylococcus aureus is a common infectious agent in e.g. sepsis, associated with both high mortality rates and severe long-term effects. The cytolytic protein α-hemolysin has repeatedly been shown to enhance the virulence of S. aureus. Combined with an unhindered spread of multi drug-resistant strains, this has triggered research into novel anti virulence (i.e. anti α-hemolysin) drugs. Their functionality will depend on our ability to identify infections that might be alleviated by such. We therefore saw a need for detection methods that could identify individuals suffering from S. aureus infections where α-hemolysin was a major determinant. Molecular imprinted polymers were subsequently prepared on gold coated sensor chips. Used in combination with a surface plasmon resonance biosensor, α-hemolysin could therethrough be quantified from septic blood samples (n = 9), without pre-culturing of the infectious agent. The biosensor recognized α-hemolysin with high affinity (KD = 2.75 x 10-7 M) and demonstrated a statistically significant difference (p < 0.0001) between the α-hemolysin response and potential sample contaminants. The detection scheme proved equally good, or better, when compared to antibody-based detection methods. This novel detection scheme constitutes a more rapid, economical, and user-friendly alternative to many methods currently in use. Heightening both reproducibility and sensitivity, molecular imprinting in combination with surface plasmon resonance (SPR)-technology could be a versatile new tool in clinical- and research-settings alike.
Highlights
Commonly isolated as an asymptomatic colonizer, Staphylococcus aureus is the leading cause of blood- (Pfaller et al, 1999), skin- and soft tissue-infections (Fridkin et al, 2005) worldwide
Our results suggest that molecular imprinted polymer (MIP) technology could be useful in detection of this diagnostic, prognostic and potentially predictive biomarker of sepsis
Limit of detection (LOD) values were calculated from the respective formulas (3xSD from blank measurements) and found to be 0.022 and 2.256 μM for the MIP- and NIP-chip respectively
Summary
Commonly isolated as an asymptomatic colonizer, Staphylococcus aureus is the leading cause of blood- (Pfaller et al, 1999), skin- and soft tissue-infections (Fridkin et al, 2005) worldwide. With functions ranging from modification of neutrophil responses (Thammavongsa et al, 2013) and complement activation (Rooijakkers et al, 2005) to host cell lysis (Powers et al, 2012), are secreted during S. aureus infections. The latter, a ß-barrel-forming cytotoxin termed ahemolysin (a-toxin, Hla), is heavily involved in the pathogenesis of sepsis due to e.g. its negative effect on endothelial integrity (Powers et al, 2012). We saw need for a novel detection method that could rapidly, with high sensitivity and selectivity, recognize patients suffering from an S. aureus infection where a-hemolysin is a major determinant
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