TY - JOUR ID - TI - Molecular technology for the detection of Pyoviridine gene in Pseudomonas aeruginosa isolated from burn cases AU - Eman Nassir Hussan AL‑Jesmany AU - Oruba Khalid Abbas AU - Basima QASIM Hasan AlSaadi PY - 2022 VL - 21 IS - 1 SP - 23 EP - 28 JO - Mustansiriya Medical Journal مجلة المستنصرية الطبية SN - 20701128 22274081 AB - Objectives: Using molecular technology for the purpose of confirmation of the diagnosis of Pseudomonas aeruginosa which depends on the detection of proteins like Pyoverdine gene of P. aeruginosa as diagnostic test. Materials and Methods: This work was done on 110 patients who had sustained burn injury from hospitals Baghdad, Al-Yarmouk and the Medical City Teaching Hospital during the period from October 2020 to the end of March 2021. The collected samples were cultured on different media (blood agar, MacConkey agar, nutrient agar, and Cetrimide agar) for isolation of P. aeruginosa bacteria as well as automated biochemical tests such as Api20E and VITEK 2 systems. The results showed that 76 (69%) samples had bacterial growth of P. aeruginosa. Antibiotic susceptibility testing was evaluated using the VITEK 2 compact. The Pseudomonas was detected using species-specific gene 16SrRNA gene using polymerase chain reaction (PCR) method and also detected by (PCR) using newly designed primers with a molecular size (389 bp) for pyoviridine gene. Results: This work showed that the predominant growth of burn wound infections was P. aeruginosa 76 samples (69%). Antibiotic susceptibility testing results showed the same sensitivity pattern of P. aeruginosa isolates to ceftriaxone and cephalothin (67.1%), gentamicin, piperacillin, ceftazidime, cefepime, and cefotaxime (65.7%). Resistance to imipenem, tobramycin, ticarcillin, and meropenem were (57.8%), (51.3%), (56.5%) and (55.2%) respectively, and had the highest sensitivity to amikacin (34.2%). Moreover, the highest resistance was to ciprofloxacin (69.7), and norfloxacin (71%). The results of using the 16SrRNA gene for the detection Pseudomonas give positive results (100%). Pseudomonas spp was detected by (PCR) for pyoviridine gene, revealed that 50% isolates give positive results. PCR product pyoviridine of the isolates (3) with the highest resistance to fluoroquinolones (ciprofloxacin and norfloxacin) was sent to nitrogen-based sequencing, and the sequencing results revealed the mutation presence. Conclusion: This study shows the current resistant pattern of P. aeruginosa against different classes of antibiotics and the involvement of several virulence genes in resistance mechanisms by using PCR which ultimately helps to select appropriate antibiotics useful for the treatment of many burned complicated by P. aeruginosa.

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