NOVEL METHODS FOR RAPID IDENTIFICATION OF BACTERIAL PATHOGENS

Abstract

Intro: Antimicrobial-resistant bacteria pose a major threat to the healthcare and agricultural system. Problems with appropriate treatment in these settings arise through the lack of fast identification of the disease-causing species and lengthy handling processes. Current methods for bacterial identification require culturing and often take days until a result can be returned. This not only means that patients experience a lag in their treatment but the potential administration of ineffective or broad-spectrum antibiotics also increases the likelihood for resistance to develop. Methods: We established three rapid methods for the detection of mastitis pathogens from bovine milk without the need to culture the pathogen. Findings: Using laser-assisted Rapid Evaporative Ionisation Mass Spectrometry (LA-REIMS) made it possible to detect specific biomarkers in less than 30 minutes. Interestingly, these molecules were found to originate from a pathogen- specific host-immune response as well as the bacteria themselves. In a second approach, a combination of portable PCR techniques using highly specific primers with a printed array strip enabled the detection of major mastitis pathogens within 120 minutes. Furthermore, the construction of a set of whole- cell bacterial biosensors allows for visual detection of pathogens without any equipment in less than 240 minutes. This system can also be modified to function as a targeted drug delivery system. Conclusion: The presented methods drastically reduce the time needed for the specification of disease-causing pathogens and enable an informed antibiotic administration from the start of the treatment, which can improve the quality of care and reduce the emergence of antimicrobial resistances.