Simultaneous detection using a portable multiplex PCR-dual lateral flow immunoassay for P. carotovorum subsp. brasiliense and E. coli O157:H7.

[Display omitted] • Multiplex PCR using portable PCR device can detect P. carotovorum subsp. brasiliense and E. coli O157:H7 without interference. • Multiple amplicons were visualized using antigen-attached dual LFA strip. • The method can detect up to 102 CFU/g in six agricultural products, which is a risk of bacterial soft-rot and food-borne disease in 1 h. A portable multiplex polymerase chain reaction (mPCR)-dual LFA system was developed for the simultaneous detection of P. carotovorum subsp. brasiliense and E. coli O157:H7 in fresh agricultural products. The field applicability of the system was improved using a portable PCR device and dual LFA strip, enabling rapid and sufficiently sensitive results to be obtained within 60 min. The mPCR-LFA method was validated for each bacterial pure culture and six agricultural products, with successful detection of both bacteria up to 102 CFU/g, a sufficiently sensitive level for ensuring the safety of all fresh agricultural products. The results demonstrate that the mPCR-LFA method developed in this study can identify pathogens much more conveniently in a point-of-care detection setting. [ABSTRACT FROM AUTHOR]