Rapid detection of three IMO standard marine bacteria using recombinase polymerase amplification and lateral flow assay (RPA-LFA)

The Convention on the International Maritime Organization (IMO), the presence of Escherichia coli, intestinal Enterococcus spp., and toxigenic Vibrio cholerae in the ballast water, prior to disposal, indicates the existence of microbes. However, current testing processes for controlling marine bacteria pathogens either require too much equipment or are too time-consuming. To improve the testing process, a combination of Recombinase Polymerase Amplification (RPA) and a Lateral Flow Assay (LFA) was used to detect V. cholera and two species of Enterococcus (E. faecalis and E. faecium) in ballast water in this study. These results showed the successful development of an RPA-LFA method for detecting V. cholera, E. faecalis and E. faecium. A variety of temperatures (30–50°C) and incubation durations (0–30 min) were investigated. Optimal results were attained at 37°C for 20 min, with lateral flow dipsticks detecting within 5–10 min and allowing diagnosis in roughly 1 h without advanced tools. This detection method is suitable for use in the on-site diagnosis also it has high specificity and is quick to use.