Figure 1
Figure 2
Antibiotic sensitivity of the examined samples (Escherichia coli and Klebsiella pneumoniae)_
| Antimicrobial | Resistant strains | Nonresistant strains | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| E. coli | K. pneumoniae | E. coli | K. pneumoniae | |||||||||
| R | S | I | R | S | I | R | S | I | R | S | I | |
| Ampicillin | 14 | 0 | 0 | 3 | 0 | 0 | 12 | 1 | 0 | 10 | 3 | 0 |
| Amoxicillin/clavulanic acid | 2 | 8 | 4 | 1 | 1 | 1 | 1 | 8 | 4 | 7 | 6 | 0 |
| Piperacillin/tazobactam | 5 | 8 | 1 | 2 | 0 | 1 | 2 | 11 | 0 | 7 | 6 | 0 |
| Cefoxitin | 2 | 9 | 3 | 1 | 1 | 1 | 0 | 11 | 2 | 7 | 6 | 0 |
| Ceftazidime | 14 | 0 | 0 | 3 | 0 | 0 | 0 | 13 | 0 | 7 | 6 | 0 |
| Cefepime | 14 | 0 | 0 | 3 | 0 | 0 | 0 | 13 | 0 | 7 | 6 | 0 |
| Imipenem | 0 | 13 | 1 | 0 | 3 | 0 | 0 | 13 | 0 | 7 | 6 | 0 |
| Meropenem | 0 | 13 | 0 | 0 | 3 | 0 | 0 | 13 | 0 | 7 | 6 | 0 |
| Amikacin | 0 | 14 | 0 | 1 | 2 | 0 | 0 | 13 | 0 | 7 | 6 | 0 |
| Gentamicin | 4 | 10 | 0 | 1 | 2 | 0 | 2 | 11 | 0 | 8 | 5 | 0 |
| Ciprofloxacin | 10 | 4 | 0 | 3 | 0 | 0 | 2 | 11 | 0 | 8 | 5 | 0 |
| Tigecycline | 0 | 14 | 0 | 0 | 2 | 1 | 0 | 13 | 0 | 7 | 5 | 1 |
| Nitrofurantoin | 2 | 11 | 1 | 2 | 0 | 1 | 0 | 13 | 0 | 10 | 0 | 3 |
| Trimethoprim/Sulfamethoxazole | 9 | 5 | 0 | 3 | 0 | 0 | 7 | 6 | 0 | 8 | 5 | 0 |
Results of polymerase chain reaction of genes encoding important ß-lactamases (bla) genes in selected bacteria and relevant antibiotic sensitivity
| Sample Number | Isolated bacteria | TEM variants including TEM-1 and TEM-2. | CTX-M group | R (resistant), S (sensitive), I (intermediate) | |||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| CTX-1 | CTX-2 | CTX-9 | Amp (ampicillin), Am/Cla (amoxicillin/clavulanic acid), Pip/Taz (piperacillin/tazobactam), Cefo (cefoxitin), Cefta (ceftazidime), Cefe (cefepime), Imi (imipenem), Mer (meropenem), Amk (amikacin), Gen(gentamicin), Cip (ciprofloxacin), Tig (tigecycline), Nit (nitrofurantoin), Tri/Sul (trimethoprim/sulfamethoxazole), E. coli = Escherichia coli and K. pneumoniae = Klebsiella pneumoniae. | Am/Cla | Pip/Taz | Cefo | Cefta | Cefe | Imi | Mer | Amk | Gen | Cip | Tig | Nit | Tri/Suif | |||
| 1 | E. coli | + | - | - | - | R | R | I | R | R | R | S | S | S | S | R | S | S | R |
| 2 | K. pneumoniae | + | + | - | - | R | R | I | R | R | R | S | S | S | S | R | S | I | R |
| 3 | E. coli | + | - | - | - | R | S | R | S | R | R | S | S | S | S | R | S | S | R |
| 4 | E. coli | + | - | - | - | R | I | R | S | R | R | S | S | S | S | R | S | S | S |
| 5 | K. pneumoniae | - | + | - | - | R | I | R | I | R | R | S | S | R | R | R | I | R | R |
| 6 | E. coli | + | - | - | - | R | S | S | S | R | R | S | S | S | S | R | S | S | R |
| 7 | E. coli | + | - | - | - | R | I | S | I | R | R | S | S | S | R | R | S | I | R |
| 8 | E. coli | + | - | + | - | R | S | R | S | R | R | I | S | S | R | R | S | S | S |
| 9 | E. coli | + | + | - | - | R | S | S | S | R | R | S | S | S | S | S | S | S | S |
| 10 | E. coli | + | - | - | - | R | I | S | S | R | R | S | S | S | S | R | S | R | R |
| 11 | E. coli | + | - | - | - | R | R | R | S | R | R | S | S | S | S | R | S | S | R |
| 12 | E. coli | + | - | - | - | R | S | S | S | R | R | S | S | S | S | S | S | S | R |
| 13 | E. coli | + | - | - | - | R | S | S | I | R | R | S | S | S | R | S | S | S | R |
| 14 | E. coli | + | + | — | — | R | S | S | R | R | R | S | S | S | S | R | S | S | S |
| 15 | E. coli | + | - | - | - | R | S | S | I | R | R | S | S | S | R | S | S | S | R |
| 16 | K. pneumoniae | + | + | - | - | R | S | R | S | R | R | S | S | S | S | R | S | R | R |
| 17 | E. coli | + | - | - | - | R | I | R | S | R | R | S | S | S | S | R | S | R | S |
Oligonucleotide sequences used to identify genes encoding important β-lactamase (bla) genes and random primers used for genotyping of the bacteria
| Primer name | Primer Sequence 5′–3′ | Position | Amplicon size |
|---|---|---|---|
| TEM-1 and TEM-2 | |||
| MultiTSO-T_forward | CATTTCCGTGTCGCCCTTATTC | 13–34 | 800 |
| MultiTSO-T_reverse | CGTTCATCCATAGTTGCCTGAC | 812–791 | |
| CTX-M group 1, group 2 and group 9 | |||
| MultiCTXMGp1forward | TTAGGAARTGTGCCGCTGYA | 61–80 | 688 |
| MultiCTXMGp1-2_reverse | CGATATCGTTGGTGGTRCCAT | 748–728 | |
| MultiCTXMGp2_forward | CGTTAACGGCACGATGAC | 345–362 | 404 |
| MultiCTXMGp1-2_reverse | CGATATCGTTGGTGGTRCCAT | 748–728 | |
| MultiCTXMGp9_forward | TCAAGCCTGCCGATCTGGT | 299–317 | 561 |
| CTXMGp9_reverse | TGATTCTCGCCGCTGAAG | 859–842 | |
| Random primers | |||
| OP-A1 | CAGGCCCTTC | ||
| OP-A3 | AGTCAGCCAC | ||
| OP-A4 | AATCGGGCTG | ||
| OP-A5 | AGGGGTCTTG | ||
| OP-A6 | GGTCCCTGAC | ||
| OP-A7 | GAAACGGGTG | ||
| OP-A8 | GTGACGTAGG | ||
| OP-A9 | GGGTAACGCC | ||
| OP-A10 | GTGATCGCAG | ||
| OP-B7 | GGTGACGCAG | ||
| OP-D5 | TGAGCGGACA |
Polymorphic bands of each genetic primers and percentage of polymorphism in the extended spectrum β-lactamase-producing bacterial isolates
| Primers | Total Bands | No. of Monomorphic Bands | No. Polymorphic Bands | % Monomorphic bands | % Polymorphic bands |
|---|---|---|---|---|---|
| OPA-01 | 19 | 4 | 15 | 26.3 | 73.7 |
| OPA-03 | 20 | 6 | 14 | 30.0 | 70.0 |
| OPA-04 | 20 | 5 | 15 | 25.0 | 75.0 |
| OPA-05 | 21 | 5 | 16 | 23.8 | 76.2 |
| OPA-06 | 16 | 0 | 16 | 0.00 | 100 |
| OPA-07 | 18 | 10 | 8 | 55.5 | 44.5 |
| OPA-08 | 19 | 6 | 13 | 31.5 | 68.5 |
| OPA-09 | 15 | 1 | 14 | 6.67 | 93.3 |
| OPA-10 | 22 | 4 | 18 | 18.2 | 81.8 |
| OPB-07 | 17 | 9 | 12 | 52.9 | 47.1 |
| OPD- 05 | 20 | 15 | 5 | 75.0 | 25.0 |
| Total | 207 | 65 | 142 |