Multiplex PCR for Aeromonas Identification

Stool samples were transported from the community clinics and were tested daily except on weekends, where part of each sample was suspended in ASL buffer (Qiagen, Hilden, Germany) and refrigerated at 4 °C until tested by PCR only (culturing was done from the original sample tube). Multiplex PCR for Aeromonas and other bacterial pathogens was performed as previously described [7 (link)]. Stool samples that were positive by PCR for Aeromonas were inoculated into alkaline peptone water 0.5 M NaCl with Cephalothin (10 mg/l) and incubated overnight [8 (link)] followed by sub-culturing onto SS agar plates (Hylabs, Rehovot, Israel). Presumptive Aeromonas colonies were identified by MALDI Biotyper Sirius system (Bruker Daltonics, Bremen, Germany) using the MBT IVD Library Revision software. Definite determination of species was based on whole genome sequencing as described below.