Table 1.
Primers sequences used for screening tested genes.
Fig 1.
Phenotypic detection of β-lactamase production.
A. Modified-double disc synergy test for determination of ESBL production. B. The 3-dimensional enzyme extract test for detection of AmpC production. C. Modified Hodge test for detection of carbapenemase production.
Fig 2.
PCR assay for β-lactamase enzyme genes.
Lane M was 100bp DNA marker and lane C was a negative control. A. Agarose gel electrophoresis of AmpC gene amplicons (254 bp). B. Agarose gel electrophoresis of blaCTX-M-15 gene amplicons (255 bp). C. Agarose gel electrophoresis of blaSHV-1 gene amplicons (356 bp). D. Agarose gel electrophoresis of blaTEM-1 gene amplicons (786 bp).
Fig 3.
Frequency of β-lactamase producing tested isolates by phenotypic analysis.
A. ESBL producers, B. AmpC producers, C. Carbapenemase producers. A statistically significant difference was detected between ESBL producers and carbapenemase producers among K. pneumoniae and similarly among E. coli without a significant difference between ESBL producers and AmpC producers in either type. Results were analyzed with Z-test for proportion with p<0.05*.
Fig 4.
Percentage of ESBL, AmpC β-lactamase, and carbapenemase genes among the tested isolates.
There is a statistically significant difference between different genes studied among K. pneumoniae and also among E. coli (p<0.001*). Results were analyzed with the Chi-Square test.
Fig 5.
PCR assay for carbapenemases enzymes genes.
Lane M was 100bp DNA marker and lane C was a negative control. A. Agarose gel electrophoresis of the blaNDM-1 gene amplicons (133 bp). B. Agarose gel electrophoresis of blaVIM-1 gene amplicons (105 bp). C. Agarose gel electrophoresis of blaOXA-48 gene amplicons (743 bp).
Fig 6.
Outer-membrane porin protein and gene analyses.
A. PCR detection of outer membrane porin encoding genes. Lane M: 100 bp plus DNA marker; lanes 1 and 2: amplified product of OmpC gene (1443 bp); lanes 3 and 4: amplified product of OmpF gene (1341 bp) which are porin genes in E. coli; lanes 5 and 6: amplified product of OmpK35 gene (1128bp) which is porin gene in K. pneumoniae. B. SDS-PAGE analysis of outer membrane protein of some E. coli isolates. C. SDS-PAGE analysis of outer membrane protein of some K. pneumoniae isolates. Lane M: marker protein; lane ST: Standard strain used as control.
Fig 7.
SDS-PAGE analysis of outer membrane proteins of sequenced genes.
A. SDS-PAGE analysis of outer membrane protein of E. coli isolates 7E, 36E, 27E and 38E. B. SDS-PAGE analysis of outer membrane protein of K. pneumoniae isolates 5K and 23K. Lane M: marker protein; Lane ST: Standard strain used as control.
Table 2.
Detected mutations in tested OmpF, OmpC and OmpK35 OMP genes.