Rapid Identification of Pseudomonas aeruginosa by Using Real Time PCR

Abstract

Pseudomonas aeruginosa is an aerobic Gram-negative bacterium which has emerged as one of the most problematic nosocomial pathogens.To characterizes P. aeruginosa strains that are widespread in patients in Iraq, 90 clinical samples were collected from wounds, burn, ear infection and urinary tract infection taken from three general hospitals of different areas of the region in Baghdad . Methods for isolation and identifying P. aeruginosa based upon culture methods coupled with biochemical tests, were used in this study. The results show that, the selective medium (cetrimide agar) at 42˚C aerobically had highest recovery in the isolation of P. aeruginosa strains, they were produced greenish-yellow or blue pigment colonies, catalase and oxidase was positive whereas negative for methyl red, and indole.; however, some of these methods are time consuming and may not be very accurate whereas API 20E is rapid method which performs at least 20 different biochemical tests at once, however it proved difficult to obtain additional information concerning the relationship between these strains. Molecular study for identifying p.aeruginosa include DNA extraction , than Real time assay by using powerchek P. aeruginosa Real time PCR Kit with probe, the results showed that RT-PCR has found to be rapid and more sensitive and specific in identification of P. aeruginosa, however Real time PCR Kit with probe, specific marker is recommended