TG TC CATA T T G G R:CGAAT CG AG AC AG G C C C F:AGCCC TA TT AC CCGCC A G A C R:CTGGC TA TT TC AT CATCCC T A C A C F:GGTTT GC AT TG TTTTC G G C G R:CGGAA GG TC TC CGATC T C A A F:TGGAA GG GA AA AGGTC G C G A R:TTGCC AA CA TC TTCCA C C G T F:GCGTG TT AG AT AACAC G A G G R:CATCA GT CA CC AACCG A T A C F:CGAGT CG CA TA CTGGT A G T G R:CTCTT GC TT CG CCCAT G T C T Amplicon size (bp) 396 1316 250 504 1310 307 1013 300 455 390 114 621 400 438 250 Reference4 of|GHASEMIAN et al.PCR method employing the particular primers (Table 1).19?2 Then, 1 agarose gel electrophoresis and gel staining (stain load dye (CinnaGen Co, Iran)) were conducted for the evaluation of PCR solutions.TA B L E 2 AntibioticsusceptibilitypatternsofP. aeruginosa isolates.Antibiotics Sensitive N ( ) 22 (55) 24 (60) 27 (67.5) 24 (60) 24 (60) 24 (60) 24 (60) 40 (100) Intermediate N ( ) 0 0 0 0 0 0 0 0 Resistant N ( ) 18 (45) 16 (40) 13 (32.5) 16 (40) 16 (40) 14 (35) 16 (40) 0 (0)two.six | Enterobacterial repetitive intergenic consensus (ERIC-PCR)To characterize the genetic relatedness amongst the isolates, ERICPCR was performed employing followed primers, ERIC1 5- TGTA GC A A TCC GG GA TCAC- and ERIC2 5- AGTA GT AC GG GT T G T three A A G T G GAGCG- , as described previously.13 The PCR protocol consisted three ofapre- enaturationstepat95 for5min,followedby30cycles d of 60s at 95 , 50s at 59 , and 60s at 72 . A final extension step was accomplished at 72 for ten min. PCR items were separated by electrophoresis in 1.5 agarose gels with 0.5?TBE (Tris/Boric acid/ EDTA)buffer.DNAbandswerevisualizedusingUVlightafterstainingwithsafestainloaddye.5-Nitro-1H-pyrazole-3-carbonitrile site TheGelJsoftwareversion2.0wasused to analyze ERIC patterns23 as well as the isolates using a similarity coefficient90 wereclusteredinthesamegenotypes.Inotherwords, theisolateswithequalormorethan90 similarityintheirbanding patterns were regarded the exact same ERIC form.Imipenem Meropenem Ciprofloxacin Ceftazidime Cefotaxime Gentamicin Piperacillin Colistin3.2 | Phenotypic assessment of ESBL, metallo- lactamase, and carbapenemaseWhile the ESBL activity was not detected in any with the isolates, 12 (30 ) isolates have been constructive for MBL, and 16 (40 ) isolates had carbapenemase activity.two.7 | Statistical analysisThe SPSS version 22.1-Bromo-3,4-difluoro-2-methoxybenzene web 0 (SPSS, Inc.PMID:24635174 ) was utilised to analyze the data. Pearson Chi-Square test was utilised to figure out the statistically important correlation between the existence of genes and antibiotic resistance or biofilm production. Also, p-value 0.05 was considered as a significance level. The results are presented as descriptive statistics in terms of relative frequency.3.3 | Biofilm formationAll the isolates (one hundred ) were optimistic for biofilm production. Seventeen (42.five ) isolates were sturdy biofilm producers and 14 (35 ) isolates have been moderate producers. Moreover, biofilm production was weak in 9 (22.5 ) isolates. The biofilm- roducer isolates p had higher levels of antibiotic resistance (Table 3).3.4 | ESBL and carbapenemase-related genesAmongtheESBLgenes,blaTEM, blaCTX , and blaSHV genes were constructive in 15 (37.five ), 8 (20 ), and six (15 ) isolates, respectively. MBL and Carbapenemase genes had been much less frequent and only blaVIM gene was present in the isolates (30 ), and blaIMP and blaNDM genes weren’t detected. Moreover, blaOXA-48 and blaOXA-23 genes had been identified in 7 (17.five ) and 1 (2.five ) isolates and no isolate possessed blaOXA-11 gene (Table 4). The co-occurrence of distinct kinds of -lactamase was noticed in 15 isolates along with the facts are shown in Table 5.three | R E S U LT S 3.1.
