Molecular identification and detection of virulence genes among Pseudomonas aeruginosa isolated from different infectious origins
,
Abstract
Background and Objectives: Pseudomonas aeruginosa possesses a variety of virulence factors that may contribute to its pathogenicity. The aim of this study was to evaluate oprI, oprL and toxA genes for PCR identification of clinical P. aeruginosa. In order to find out any relation between special virulence factors and special manifestation of P. aeruginosa infections, we detected virulence factors among these isolates by PCR. Ribotyping was used to evaluate the clonal relationship between strains with the same genetic patterns of the genes studied.
Materials and Methods: In this study, 268 isolates of P. aeruginosa were recovered from burn, wound and pulmonary tract infections. The prevalence of oprI, oprL, toxA, lasB, exoS and nan1 genes was determined by PCR. One hundred and four isolates were selected randomly to investigate clonal diversity of the isolates with ribotyping using SmaI.
Results and Conclusions: All P. aeruginosa isolates in this study carried oprI, oprL and lasB genes. Difference between exoS prevalence in isolates from pulmonary tract and burn isolates was statistically significant. Prevalence of nan1 and toxA gene was significantly higher in pulmonary tract and burn isolates, respectively. Ribotyping showed that most of the isolates (87%) belonged to clone A and B.Detection of oprI, oprL and toxA genes by PCR is recommended for molecular identification of P. aeruginosa. Determination of different virulence genes of P. aeruginosa isolates suggests that they are associated with different levels of intrinsic virulence and pathogenicity. Ribotyping showed that strains with the same genetic patterns of the genes do not necessarily have similar ribotype patterns.
Fegan M, Francis P, Hayward AC, Davis GH, Furest JA, Phenotypic conversion of Pseudomonas aeruginosa in cystic fibrosis. J Clin Microbiol 1990; 28: 1143-1146.
Yetkin G, Otlu B, Cicek A,Kuzucu C, Durmaz R, Clinical, microbiologic, and epidemiologic characteristics of Pseudomonas aeruginosa infections in a university hospital, Malatya, Turkey. Am J Infect Control 2006; 34: 188-192.
Khan AA, Cerniglia CE, Detection of Pseudomonas aeruginosa from clinical and environmental samples by amplification of the exotoxin A gene using PCR. Appl Environ Microbiol 1994; 60: 3739-3745.
de Vos D, Lim A, Pirnay JP, Struelens M, Vandenveld C, Duinslaeger L, et al. Direct detection and identification of Pseudomonas aeruginosa in clinical samples such as skin biopsy specimens and expectorations by multiplex PCR based on two outer membrane genes, oprI and oprL. J Clin Microbiol 1997; 35: 1295-1299.
Masuda N, Sakagawa E, Ohya S. Outer membrane proteins responsible for multiple drug resistance in Pseudomonas aeruginosa. Antimicrob Agents Chemother 1995; 39: 645-649.
Nikaido, H. Prevention of drug access to bacterial targets: permeability barriers and active efflux. Science 1994; 264: 382–388.
Van Delden C, Iglewski BH. Cell-to-cell signaling and Pseudomonas aeruginosa infections. Emerg Infect Dis 1998; 4: 551-560.
Yahr TL, Hovey AK, Kulich SM, Frank DW, Transcriptional analysis of the Pseudomonas aeruginosa exoenzyme S structural gene. J Bacteriol 1995; 177:1169-1178.
Fleiszig SM, Wiener-kronish JP, Miyazaki H, Valias V, Mostov KE, Kanada D, et al. Pseudomonas aeruginosa- mediated cytotoxicity and invasion correlate with distinct genotypes at the loci encoding exoenzyme S. Infect Immun 1997; 65: 579-586.
Jaffar-Bandjee MC, Lazdunski A, Bally M, Carrere J, Chazalette JP, Galabert C, Production of elastase, exotoxin A, and alkaline protease in sputa during pulmonary tract exacerbation of cystic fibrosis in patients chronically infected by Pseudomonas aeruginosa. J Clin Microbiol 1995; 33: 924-929.
Bryan R, Kube D, Perez A, Davis P, Prince A, Overproduction of the CFTR R domain leads to increased levels of asialoGM1 and increased Pseudomonas aeruginosa binding by epithelial cells. Am J Respir Cell Mol Bio 1998; 19: 269-277.
Grattard F, Pozzetto B, Ros A, Gaudin OG.Differentiation of Pseudomonas aeruginosa strains by ribotyping: high discriminatory power by using a single restriction endonuclease. J Med Microbiol 1994; 40:275-281.
Lennette EH, Balows A, Hausler WJ, Shadom HJ, Manual of clinical Microbiology. 4th ed. Washington D.C. 1985.
Shahcheraghi F, Nikbin VS, Feizabadi MM. Prevalence of ESBLs genes among multidrug-resistant isolates of Pseudomonas aeruginosa isolated from patients in Tehran. Microb Drug Resist 2009; 15: 37-39.
Lanotte P, Mereghetti L, Dartiguelongue N, Rastegar- Lari A, Gouden A, Quentin R, Genetic features of Pseudomonas aeruginosa isolates cystic fibrosis patients compared with those of isolates from other origins. J Med Microbiol 2004; 53: 73-81.
Regnault B, Grimont F, Grimont PAD. Universal ribotyping method using a chemically labeled oligonucleotide probe mixture. Res Microbiol 1997;148: 649-659.
Qin X, Emerson J, Stapp J, Stapp L, Abe P, Burns L, Use of real-time PCR with multiple targets to identify Pseudomonas aeruginosa and other nonfermenting gram-negative bacilli from patients with cystic fibrosis. J Clin Microbiol 2003; 4: 4312-4317.
Lavenir R, Jocktane D, Laurent F, Nazaret S, Cournoyer B. Improved reliability of Pseudomons aeruginosa PCR detection by the use of the specific ecfx gene target. J Microbiol Methods 2007; 70: 20-29.
Vasil ML, Chamberlain C, Grant CCR. Molecular studies of Pseudomonas exotoxin A gene. Infect Immun 1986; 52: 538-548.
Lomholt JA, Poulsen K, Kilian M. Epidemic population structure of Pseudomons aeruginosa: evidence for a clone that is pathogenic to the eye and that has a distinct combination of virulence factor. Infect Immun 2001; 69:6284-6295.
Nicas TI, Iglewski BH. Production of elastase and other exoproducts by environmental isolates of Pseudomonas aeruginosa. J Clin Microbiol 1986; 23: 967-969.
Cowell BA, Twining SS, Hobden JA, Kwong MSF, Fleiszig SM, Mutation of lasA and lasB reduces Pseudomonas aeruginosa invasion of epithelial cells. Microbiology 2003; 149: 2291-2299.
Winstanley C, Kaye SB, Neal TJ, Miksch S, Hart CA. Genotypic and phenotypic characterization of Pseudomonas aeruginosa isolates associated with ulcerative keratitis. J Med Microbiol 2005; 54: 519-526.
Feltman H, Schulert G, Khan S, Jain M, Peterson L, Hauser AR, Prevalence of type III secretion genes in clinical and environmental isolates of Pseudomonas aeruginosa. Microbiology 2001; 147: 2659-2669.
Schulert GS, Feltman H, Rabin SDP, Martin CG, Battle SE, Rello J, et al. Secretion of the toxin ExoU is a marker for highly virulent Pseudomonas aeruginosa isolates obtained from patients with hospital-acquired pneumonia. J Infect Dis 2003; 188: 1695-1706.
Botes J, Williamson G, Sinickas V, Gürtler V.Genomic typing of Pseudomonas aeruginosa isolates by comparison of Riboprinting and PFGE: correlation of experimental results with those predicted from the complete genome sequence of isolate PAO1. J Microbiol Methods 2003; 55: 231-240.
| Files | ||
| Issue | Vol 4 No 3 (2012) | |
| Section | Articles | |
| Keywords | ||
| Pseudomonas aeruginosa ribotyping virulence factors | ||
| Rights and permissions | |
|
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
