Identification of Clinical Pseudomonas spp. by VITEK 2 Compact System and Species-specific Polymerase Chain Reaction Assay for Identification of Pseudomonas aeruginosa

Abstract

The objective of this study was to isolate, identify, and diagnose Pseudomonas spp. from different clinical sources in Baghdad, Iraq. VITEK 2 compact system identification gram-negative bacteria (ID gNB) cards were used to confirm the identification. Polymerase chain reaction (PCR) technique and sequencing were used for recognition of the 16S rDNA gene, by two pairs of primers, universal primers (930 bp fragments) for recognition of Pseudomonas spp., and Pseudomonas aeruginosa specific species (PASS) primers (956 bp fragments) for differentiation of P. aeruginosa from other species. Amplified PCR products of PASS primers were sent for DNA Sanger sequencing to Macrogen Company, Seoul, Korea; data were compared with the database using the Basic Local Alignment Search Tool (BLAST). Ninety-two Pseudomonas spp., including 86 isolates of P. aeruginosa and 1, 2, 3 isolates of Pseudomonas luteola, Pseudomonas putida, and Pseudomonas fluorescens, respectively, were obtained using VITEK 2 compact system ID gNB cards with a percentage of identification ranging from 91 to 99%. Gene amplification and sequencing results confirm identification ranging from 99 to 100%. After sequencing analysis, eleventh of the P. aeruginosa isolates had been submitted in GenBank National Centre for Biotechnology Information (NCBI) under accession numbers MN630696–MN630707. It was observed that phenotypic tests supported by PCR techniques have enabled to conduct a detailed characterization of Pseudomonas bacteria isolates.