Background: P. aeruginosa is an important nosocomial pathogen with increasing resistance to antibiotics. Objective: This study
was performed to evaluate the genetic relatedness of MDR clinical isolates of P. aeruginosa.

Method: A total of 1000 samples were analysed in the study. Antibiotic resistance profiles of the isolates were determined using
Kirby Bauer disk diffusion method. Polymerase chain reaction (PCR) and sequencing were simultaneously used to detect the
consensus region of 16S rRNA. Genetic relatedness of the isolates was determined using restriction patterns from ALU 1 digest
and random amplified polymorphic DNA.

Results: Out of the 192 P. aeruginosa isolates recovered, 136 (78.83%) were multidrug resistant. Sequence analysis of the confirmed
isolates (80.68%) revealed that all the isolates shared homology with each other and also showed sequence similarity to
known strains of P. aeruginosa (ATCC 27853; KT 315654; KU 321274 and KT894767). The PCR-Restriction fragment length
polymorphism (PCR-RFLP) analysis revealed that there was a lot of genetic relatedness among the isolates. The RFLP finger
printing technique detected seven distinct RFLP types among the isolates.

Conclusions: Thus, study shows that there is high prevalence of MDRPA and high degree of genetic relatedness among the
MDRPA isolates circulating in Nsukka area.

Keywords: MDRPA; PCR-RFLP; RAPD; Sequencing; 16S rRNA gene.