Molecular search of some antibiotic-borne genes of the clinical Pseudomonas aeruginosa plasmid

Abstract

Clinical samples which included wounds, burns, ear infections, urinary tract infections (UTI), and skin from 300 patient from a different hospital in Thi-Qar province, and 100 skin and stool samples from (healthy people) as control, for investigating the presence of pseudomonas spp. Where carried out through October 2019 till January 2020. The samples were identified based on the morphological , microscopical characteristics of the colonies and biochemical tests, then confirmed by API-20 NE, VITEK2- compact system and the molecular detection based on the 16SrRNA gene, final diagnosis showed that 60 isolates belong to target bacteria were distributed as 30(50%) of burns, 5(8.3) isolates of wounds, 7(11.6) isolates of ear infection, 4(6.6) isolates (sputum),4(6.6)isolates of urinarytract infection, and (8 (13.3),2(3.3)) isolates stool samples and skin swabs respectively, as control samples). All isolates were tested for susceptibility test toward 13 antibiotics using the disc diffusion method, the results showed a high resistance among isolates against Tetracycline, Gentamycin, Amikacine. moderate resistance against Cefepime, Ceftazidime, and Imipenem. And low resistance towards Meropenem, and Ciprofloxacin. The results of antibiotic-resistance genes was diagnostic by Polymerase chain reaction (PCR) amplify based on specific sequences for (MBLs genes)bla VIM ,bla IMP, and bla NDM genes, showed that 26 (43.3%) of isolates have bla VIM gene, 16 (26.6%) of isolates have bla IMP gene, and 11 (18.3%)of isolates have bla NDM gene. while the results of (AME-genes) indicated that 30 (50%) of isolate have aac (6)-I gene and 8(13.3%) of isolate have aph (3)-VI gene. A comparison is made between the presence of genes on the plasmid or the chromosome,the results was increased the antibiotic- resistance genes on the plasmid. DNA partial sequencing was done for four isolates revealed some variants in different samples while some isolates showed 100% similarity with the referencing genome.