Table 1.
Sequence of oligonucleotide primers used for PCR amplification of Klebsiella spp., ESBLs encoding genes and biofilm genes.
| Target gene | Primer sequence (5ʹ–3ʹ) | Amplicon size (bp) | Amplification conditions | |
|---|---|---|---|---|
| Klebsiella spp. | gyrA |
F: CGC GTA CTA TAC GCC ATG AAC GTA R: ACC GTT GAT CAC TTC GGT CAGG |
441 | Initial denaturation of 5 min at 94 °C followed by 35 cycles, denaturation at 94 °C for 30 s, annealing at 55 °C for 45 s, extension at 72 °C for 45 s, and final extension at 72 °C for 10 min |
| K. pneumoniae | magA |
F: ATT TGA AGA GGT TGC AAA CGAT R: TTC ACT CTG AAG TTT TCT TGT GTTC |
130 | Initial denaturation of 5 min at 94 °C followed by 35 cycles, denaturation at 94 °C for 30 s, annealing at 55 °C for 30 s, extension at 72 °C for 40 s, and final extension at 72 °C for 10 min |
| ESBLs encoding genes | bla TEM |
F: CGC CGC ATA CAC TAT TCT CAG AAT GA R: ACG CTC ACC GGC TCC AGA TTT AT |
445 | Initial denaturation of 5 min at 94 °C followed by 30 cycles, denaturation at 94 °C for 1 min, annealing at 61 °C for 1 min, extension at 72 °C for 1 min, and final extension at 72 °C for 5 min |
| bla SHV |
F: CTT TAT CGG CCC TCA CTCAA R: AGG TGC TCA TCA TGG GAA AG |
237 | ||
| bla CTX-M |
F: ATG TGC AGY ACC AGTAAR GTK ATG GC R: TGG GTR AAR TAR GTS ACC AGA AYC AGC GG |
593 | ||
| bla OXA-1 |
F: ACA CAA TAC ATA TCA ACT TCG C R: AGT GTG TTT AGA ATG GTG ATC |
813 | ||
| biofilm genes | LuxS |
F: GCC GTT GTT AGA TAG TTT CACAG R: CAG TTC GTC GTT GCT GTT GATG |
447 | Initial denaturation of 2 min at 95 °C followed by 35 cycles, denaturation at 95 °C for 20 s, annealing at 53 °C for 30 s, extension at 72 °C for 45 s, and final extension at 72 °C for 7 min |
| Uge |
F: TCT TCA CGC CTT CCT TCA CT R: GAT CAT CCG GTC TCC CTG TA |
535 | ||
| mrkD | F: CCACCAACTATTCCCTCGAA R: ATGGAACCCACATCGACATT | 226 |