Application of Modern Methods for the Determination of Antibiotic Resistance

The increasing occurrence of antibiotic resistance is one of the major problems of the 21st century. The occur-rence of bacterial strains resistant to antibiotics subse-quently narrows the spectrum of suitable antibiotics usable for the treatment of common bacterial infections or for the prevention of their occurrence, e.g., in surgery. Wastewater treatment plants, hospitals, and also the food chain belong to the hotspots, where the emergence and spread of new or existing strains of antibiotic resistant bacteria and antibiotic resistance genes occur most fre-quently. Phenotypic culture methods are routinely used in laboratories to determine antibiotic resistance, but they are laborious and time-consuming and the interpretation of exact results is also difficult. For this reason, faster alter-natives for the detection of antibiotic resistant bacteria or even antibiotic resistance genes are sought. Such an exam-ple of an alternative method for the detection of antibiotic resistant bacteria is the use of the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry phenotypic method to identify the beta-lactamase produc-ers. Genotype methods provide faster analysis and, at the same time, more accurate detection. Antibiotic resistance genes can be directly detected and quantified by polymer-ase chain reaction. Microarrays can be used to further speed up and increase the specificity of PCR amplicons detection. Massive parallel methods provide comprehen-sive information on the resistoma of the specific environ-ment. They facilitate sequencing of individual DNA mole-cules or amplicons to detect determinants of antibiotic resistance. Massive parallel methods have the potential to replace conventional pathogen characterization and allow the detection of all microorganisms in a sample (including difficult-to-cultivate or noncultivable microorganisms).