Rapid and simultaneous analysis of twelve virulence factor genes by a microfluidic-CFPA chip for identifying diarrheagenic Escherichia coli.

Diarrheagenic Escherichia coli as an enteropathogen has caused serious public safety problems, especially in children. A fast, easy-to-use, high-throughput method is urgently needed for diarrheagenic Escherichia coli strain identification and monitoring. In this study, we developed a portable microfluidic device based on circular fluorescent probe-mediated isothermal nucleic acid amplification for accurate and rapid detection of twelve virulence factor genes of five diarrheagenic Escherichia coli strains for point-of-care testing. This microfluidic system showed excellent performance in identifying five diarrheagenic Escherichia coli strains within 60 min, when applied in 67 clinical samples collected from hospitals for identifying diarrheagenic Escherichia coli, with a good clinical sensitivity (96.9%), specificity (97.1%) and stability (CV < 5.0%). This integrated microfluidic system is a promising diagnostic tool for applications in the identification and monitoring of diarrheagenic Escherichia coli epidemics worldwide, particularly in developing countries.