title: Detection of algD, oprL and exoA Genes by New Specific Primers as an Efficient, Rapid and Accurate Procedure for Direct Diagnosis of Pseudomonas aeruginosa Strains in Clinical Samples.
abstract: BACKGROUND
  Pseudomonas aeruginosa is an opportunistic pathogen that infects patients with cystic fibrosis, burning wounds, ophthalmic traumas and immunodeficiency.
  
  
  OBJECTIVE
  The aim of the present study was to compare the efficiency of newly designed primer sets with some previously published primers for PCR detection of exoA, oprL and algD genes from P. aeruginosa.
  
  
  METHODS
  A total of 150 clinical specimens were inoculated into the routine and selective culture media for P. aeruginosa isolation. Specific primers were designed by bioinformatics analysis for detection of the virulence genes determinants, exoA, oprL and algD. The sequences of these three genes were obtained from NCBI and multiple alignments were performed to find the conserved sequences of each gene to design the primers. Multiple alignment and primer design steps were carried out by the AlleleID software, version 7.0.
  
  
  RESULTS
  Microbiological culture methods resulted 70 P. aeruginosa strains isolated from 70 of the 150 clinical specimens. The results of the PCR assay using the newly designed exoA, oprL and algD primer sets were positive in 68, 70 and 69 clinical samples which represent 97.2%, 100% and 98% sensitivity for each primer set, respectively. The PCR results using previously published exoA, oprL and algD primer sets were positive only in 57, 49 and 28 specimens that correspond to 81.5%, 70% and 40% sensitivity, respectively.
  
  
  CONCLUSIONS
  The results of the present study showed that in comparison with previously published primer sets, P. aeruginosa infection can be diagnosed with higher sensitivity and specificity by the conventional PCR assay using the newly designed primers. It was also shown that the results of the PCR assay on clinical samples of severe infections became positive much earlier than the results of conventional culture method.
authors: Department of Medical Microbiology and Immunology, Faculty of Medicine, Arak University of Medical Sciences, Arak, IR Iran.
authors: Sedighe
authors: S
authors: Rashno Taee
authors: Department of Medical Microbiology and Immunology, Faculty of Medicine, Arak University of Medical Sciences, Arak, IR Iran.
authors: Behzad
authors: B
authors: Khansarinejad
authors: Department of Medical Microbiology and Immunology, Faculty of Medicine, Arak University of Medical Sciences, Arak, IR Iran.
authors: Hamid
authors: H
authors: Abtahi
authors: Department of Microbiology, Faculty of Medicine, Hamadan University of Medical Sciences, Hamadan, IR Iran.
authors: Mohammad
authors: M
authors: Najafimosleh
authors: Department of Medical Microbiology and Immunology, Faculty of Medicine, Arak University of Medical Sciences, Arak, IR Iran ; Molecular Research Center, Faculty of Medicine, Arak University of Medical Sciences, Arak, IR Iran.
authors: Ehsanollah
authors: E
authors: Ghaznavi-Rad
keywords: Clinical Samples, Detection, PCR; Pseudomonas aeruginosa
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