Skepple, Andre (2013) The development of a rapid immunoassay detection system for clinical and veterinary isolates of methicillin resistant 'Staphylococcus aureus'. (MSc(R) thesis), Kingston University, .

Abstract

The importance of Staphylococcus aureus and MRSA to the healthcare industry (both animal and human) is well understood. As such there is a need for rapid, sensitive and specific detection of MRSA in the clinical setting. Present gold¬standard procedures in detecting MRSA bacteria are both timely and immobile. MRSA is not only a problem in human health it is also a concern in the veterinary setting, with this organism being implemented in such veterinary diseases such as bovine mastitis. In-addition to this, the emergence and sporadic transmission of new infections such as MRSA ST398 from animals to humans has become a concern. The Vantix™ immunoassay-based biosensor technology facilitates a significant reduction in both antigen and antibody detection times using sensitive, and versatile biosensors within a portable transducer system connected to a personal computer. The primary aim of this study was to investigate if Vantix biosensors could be developed and utilised towards the rapid detection of MRSA and S. aureus. A panel of 10 monoclonal mouse anti-PBP2a antibodies (mAb's) underwent immunogenic analysis using enzyme-linked immunosorbent assay (ELISA) and SDS-PAGE Western blotting immunoassay testing, to which a pair of highly performing capture and detection anti-PBP2a mAb's, along with a polyclonal rabbit anti-S. aureus antibody was selected for Vantix biosensor immunoassay development. These biosensors were able to successfully detect clinical isolates of MRSA along with veterinary strains of MRSA ST398 using Vantix biosensors coated with the aforementioned annbodles. These biosensors were also able to detect the presence of the bacteria in artificial sweat and saliva media. Data had shown promising results with the system still fully able to detect the bacteria, which gives great promise for the Vantix biosensor platform to be further developed as a MRSA Point-of-Care (POC) screening system in the future.

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