The relation between mvat, Cupa and Cupb genes with gene expression in Isolated Pseudomonas Aeruginosa from Patients

This study aimed to study the relationship between CupA, CupB and MvaT genes in P.aeruginosa clinical isolates. Hundred fifty samples were collected, hundred one of them were diagnosed as P.aeruginosa were collected from patients with different diseases, wounds, burns, diabetic foot ulcers, urinary tract infection, keratitis, external otitis in Baghdad hospital (Al-Yarmuk General Teaching Hospital). Participants were drawn from a variety of age groups and genders. The collected samples were cultured on different selective and differential media and tested biochemically in order to identify P.aeruginosa, the identification of P. aeruginosa confirmed by using the systems API 20E, the results showed that the sensitivity for API20NE test was (80.1%). Conventional PCR has been applied for detect CupA, CupB and MvaT genes for 5 isolates, then the presence of these genes was confirmed by electrophoresis and appearance of DNA band in the agarose gel for three genes the results was positive 5/5 (100%), then sequencing was performed for the 5 isolates which gave positive results of CupA, CupB, MvaT genes and the alignment in NCBI showed identities of CupA gene 98%, CupB gene 96% and MvaT gene 100% with P. aeruginosa. Nine antibiotics were used to evaluate the antibiotic sensitivity of all isolates. The most isolates were sensitive to Levofloxacin (94%), Ticarcillin-clavulanic acid (Timinim) (88.1%), Gentamicin (82.1%) and to both Pipracillin and Imipenem (80. 1%), while the isolates appeared resistant to other antimicrobials represented by (72.2%) to Augmentin, (33.6%) to Ceftazidime, (37.6%) to Colistin, and (27.7%) to Ciprofloxacin. Levofloxacin represents the most effective antibiotic against P. aeruginosa isolates. By Real Time PCR the gene expression of CupB, MvaT and RpoD (housekeeping gene) genes were conducted for 16 isolates. It was found that the mvaT gene have the highest value of gene expression fold which was (3.86), while the CupB gene was 0.01, and the RpoD was 1.02. This study concluded that; CupB gene which have a role in biofilm formation repressed by MvaT global regulator gene and despite this, these isolates were pathogens and had resistance for some antibiotics.