جداسازى و شناسايى سريع سودوموناس آئروژينوزا از طريق PCR

Isolation and rapid detection Pseudomonas aeruginosa by PCR

Pseudomonas aeruginosa is a gram negative, opportunistic, and pathogenic bacterium that Its ubiquitous presence in the environment is a result of it sability to adapt to various adverse environmental conditions. The most serious infections caused by P aeruginosa in humans range from acute infections like endocarditis, meningitis, and septicemia to chronic lung infections in cystic fibrosis patients. Many infections can be attributed to a general immune suppression such as in AIDS patients, bum victims and neutropenic patients undergoing chemotherapy. Pseudomonas aeruginosa product various of proteins toxic and virulence factors, of which exotoxin A is the most toxic. ETA by ribosylation of elongation factor 2 inhibit protein synthesis in the eukaryotes and cell death. Therefore, accurately identifying P aeruginosa in early time is beneficial to control due infection and reduce mortality variant methods are for detection Pseudomonas aeruginosa but PCR method detected Pseudomonas aeruginosa befor proteins and virulence factors expression. This method rapid, sensitive, proprietary, inexpensive and the different than to Biochemical and serological tests. In this study, a pair of specific primers were designed in the conservative region of ETA gene by the method of bioin-formatics analysis. Pseudomonas aeruginosa sample seventy were isolated from burn patients bedridden at Moosavi Hospital, Zanjan, Iran and Pseudomonas aeruginosa species were identified by Biochemical tests. Bacteria genomic DNA was extracted and the detection by method PCR for PA was successfully developed. The specificity of the PCR was determined by analyzing 5 different strains. DNA of Staphylococcus aureus, Salmonella paratify A, Vibrio cholera, Shigella and Escherichia coll was taken as negative control. The results showed that only 4 P aeruginosa strains isolated from bum patients without ETA gen and 66 isolated Pseudomonas aeruginosa strains were asset ETA gen that specificity test determine 94.3 percent. The results sensitivity determine showed for five strain bacteria negative.