One-step species-specific high resolution melting analysis for nosocomial bacteria detection

Abstract

Nosocomial infections are a major public health concern worldwide. Early and accurate identification of nosocomial pathogens which are often multidrug resistant is crucial for prompt treatment. Hence, an alternative real-time polymerase chain reaction coupled with high resolution melting-curve analysis (HRMA) was developed for identification of five nosocomial bacteria. This assay targets species-specific regions of each nosocomial bacteria and produced five distinct melt curves with each representing a particular bacterial species. The melting curves were characterized by peaks of 78.8±0.2°C for Acinetobacter baumannii, 82.7±0.2°C for Escherichia coli, 86.3±0.3°C for Klebsiella pneumoniae, 88.8±0.2°C for Pseudomonas aeruginosa and 74.6±02°C for methicillin-resistant Staphylococcus aureus. The assay was able to specifically detect the five bacterial species with an overall detection limit of 2×10−2ng/μL. In conclusion, the HRM assay developed is a simple and rapid method for identification of the selected nosocomial pathogens.