Development and Application of SRCA for Rapid Detection of Pseudomonas aeruginosa in Bottled Drinking Water

Pseudomonas aeruginosa is a significant waterborne pathogen that poses substantial public health risks, particularly in bottled drinking water. Traditional methods for detecting P. aeruginosa are often time-consuming, expensive, and require complex equipment. This study developed a novel Saltatory Rolling Circle Amplification (SRCA) method for the rapid, sensitive, and specific detection of P. aeruginosa in bottled drinking water. The technique utilizes a simple water bath for isothermal DNA amplification and incorporates SYBR Green I for visual colorimetric detection, eliminating the need for complex post-amplification processes such as gel electrophoresis. The SRCA assay demonstrated high sensitivity with a detection limit of 9.1 × 10¹ CFU/mL and exhibited excellent specificity, as amplification products were only detected in samples containing P. aeruginosa DNA. A clear colorimetric response allowed straightforward interpretation, with positive samples exhibiting distinct green fluorescence. Evaluation using 200 artificially contaminated samples revealed that the SRCA method achieved 100.00% sensitivity, 98.90% specificity, and 99.00% accuracy compared to conventional microbiological culture. This SRCA-based assay therefore represents a promising tool for rapid, cost-effective, and on-site detection of P. aeruginosa in drinking water, with potential applications for enhancing public health safety in both industrial and field environments.