TABLE 1.

PCR primers and conditions

GenePrimers (forward, reverse)PCR conditionsaReference
manB5′-CAT AAY CCG ATG GAC TAC AAC G-3′, 5′-ACC AGC AGC CAC GGG ATC AT-3′95°C/9.5 min (1 cycle); 95°C/45 s, TD 55-45°C/45 s, 72°C/60 s (40 cycles); 72°C/7 min (1 cycle)32
mdh5′-GAT GAA AGT CGC AGT CCT CG-3′, 5′-TAT CCA GCA TAG CGT CCA GC-3′95°C/9.5 min (1 cycle); 95°C/45 s, TD 58-48°C/45 s, 72°C/60 s (40 cycles), 72°C/7 min (1 cycle)32
fimA5′-TCA GGG GAG AAA CAG AAA ACT AAT-3′, 5′-TCC CCG ATA GCC TCT TCC-3′95°C/9.5 min (1 cycle); 95°C/45 s, 57°C/45 s, 72°C/60 s (35 cycles); 72°C for 7 min (1 cycle)32
blaCMY-25′-TGG CCA GAA CTG ACA GGC AAA-3′, 5′-TTT CTC CTG AAC GTG GCT GGC-3′95°C/9.5 min (1 cycle); 95°C/45 s, 60°C/45 s, 72°C/60 s (40 cycles); 72°C/7 min (1 cycle)26
ampC5′-AAC ACA CTG ATT GCG TCT GAC-3′, 5′-CTG GGC CTC ATC GTC AGT TA-3′95°C/9.5 min (1 cycle); 95°C/45 s, 60°C/45 s, 72°C/60 s (40 cycles); 72°C/7 min (1 cycle)26
floR5′-CTG AGG GTG TCG TCA TCT AC-3′, 5′-GCT CCG ACA ATG CTG ACT AT-3′95°C/9.5 min (1 cycle); 95°C/30 s, 55°C/60 s, 72°C/60 s (40 cycles); 72°C/7 min (1 cycle)7
  • a TD, touchdown PCR; over the first 20 cycles annealing temperature is decreased by 0.5°C/cycle, followed by 20 cycles at the lower annealing temperature (i.e., temperature reached after the last TD cycle).