Genetic Detection of Antibiotic-Resistant Bacteria Using PCR and DNA Sequencing Techniques

Antibiotic resistance poses a global threat to human health and well-being. Conventional approaches to detecting resistance, such as culture and sensitivity testing, are time-consuming and do not always correlate with emerging resistance mechanisms. Molecular detection methods, including polymerase chain reaction (PCR) and DNA sequencing, have been integrated into routine diagnostic workflows for identifying clinically significant resistance and surveillance of outbreak situations. These approaches are complementary to culture-based methods and allow detection of undetectable resistance markers, confirming resistance predicted by phenotypic testing, and determination of variant alleles that modify resistance profiles. Detection workflows commence with sample processing and conventional or quantitative real-time PCR, followed by Sanger or next-generation sequencing. Variant-calling pipelines paired with publicly available resistance and annotation databases provide rapid information on known and novel mutations of clinical concern. Parallel phylogenetic analyses inform strain lineage assignments and dissemination routes, supporting surveillance and outbreak investigation.