Volume 7, Issue 4 (10-2019)                   JoMMID 2019, 7(4): 127-131 | Back to browse issues page

XML Print

Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
Khalili-Samani M, Barati M, Mirmohammadsadegh N, Amin M, Samadikuchaksaraei A. Detection of Mutations of Antimutator Gene pfpI in Pseudomonas aeruginosa Species Isolated from Burn Patients in Tehran, Iran. JoMMID 2019; 7 (4) :127-131
URL: http://jommid.pasteur.ac.ir/article-1-166-en.html
Detection of Mutations of Antimutator Gene pfpI in Pseudomonas aeruginosa Species Isolated from Burn Patients in Tehran, Iran
Maryam Khalili-Samani , Mahmood Barati , Navid Mirmohammadsadegh , Mohsen Amin * , Ali Samadikuchaksaraei
Department of Drug and Food Control, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran
Abstract:   (4652 Views)
Introduction: Pseudomonas aeruginosa is an opportunistic pathogen of clinical importance, particularly in immunocompromised and burn patients. This bacterium is becoming resistant to many antibiotics via intrinsic or acquired mechanisms. Mutations in anti-mutator genes, such as pfpI, can be a potential intrinsic mechanism of antibiotic resistance. This study aimed to evaluate the possible effects of mutations of this gene on coding proteins of multi-drug resistant P. aeruginosa isolates. Methods: The antibiotic resistance pattern of 50 P. aeruginosa isolates against 9 anti-pseudomonas antibiotics was determined by the disk diffusion method. PCR, followed by sequencing, detected the mutations in the pfpI gene. The retrieved sequences were translated to the corresponding amino acid sequences using an online protein database. The amino acid sequences in mutated isolates were compared with the reference sequence using a multiple alignment method. Results: Out of 50 isolates, 43 (86%) were resistant to all antibiotics. Sequencing and multiple alignment analyses showed that amino acids in positions 21, 24, and 57 of pfpI gene were changed in resistant isolates, and all these mutations were observed in each isolate. Homology modeling showed that these amino acids were part of a cleft on the protease. The other point mutations resulted in amino acid changes were in positions 67, 83, and 165. Conclusion: The data obtained in this study showed that the pfpI gene of P. aeruginosa might have a significant effect on response to antibiotics. Further epidemiologic and comprehensive studies are required to confirm these findings. 
Keywords: Burns, Pseudomonas aeruginosa, Drug Resistance, Amino Acid Sequence, Point Mutation
Full-Text [PDF 451 kb]   (641 Downloads)    
Type of Study: Original article | Subject: Anti-microbial agents, resistance and treatment protocols
Received: 2018/06/14 | Accepted: 2020/03/12 | Published: 2020/03/12
References
1. Church D, Elsayed S, Reid O, Winston B, Lindsay R. Burn wound infections. Clin Microbial Rev. 2006; 19 (2): 403-34. [DOI:10.1128/CMR.19.2.403-434.2006]
2. Santucci SG, Gobara S, Santos CR, Fontana C, Levin AS. Infections in a burn intensive care unit: experience of seven years. J Hosp Infect. 2003; 53: 6-13. [DOI:10.1053/jhin.2002.1340]
3. Pirnay JP, De Vos D, Cochez C, Bilocq F, Pirson J, Struelens M, et al. Molecular epidemiology of Pseudomonas aeroginosa colonization in a burn unit: persistence of multidrug-resistant clone and a silver sulfadiazine-resistant clone. J Clin Microbial. 2003; 41 (3): 1192-202. [DOI:10.1128/JCM.41.3.1192-1202.2003]
4. Rodríguez-Rojas A, Bla'zquez J. The Pseudomonas aeruginosa pfpI gene plays an antimutator role and provides general stress protection. J Bacteriol. 2009: 844-850. [DOI:10.1128/JB.01081-08]
5. Forbes BA, Sahm DF, Weissfeld AS. Diagnostic microbiology. Baily & Scott. 2007; 11th Ed. St. Louis, Mosb Inc: 389-397.
6. Clinical and Laboratory Standards Institute. Performance Standards for Antimicrobial Susceptibility Testing. (2012) 12th Informational Supplement. CLSI document M100-S12, vol. 22, No.1, Pennsylvania, USA.
7. Blazquez J. Hypermutation as a factor contributing to the acquisition of antimicrobial resistance. Clin Infect Dis. 2003; 37: 1201-9. [DOI:10.1086/378810]
8. Oliver A, Canton RP, Campo F, Baquero F, and Blazquez J. High frequency of hypermutable Pseudomonas aeruginosa in cystic fibrosis lung infection. Science. 2000; 288: 1251-4. [DOI:10.1126/science.288.5469.1251]
9. Oliver A, Levin BR, Juan C, Baquero F, and Blazquez J. Hypermutation and the preexistence of antibiotic-resistant Pseudomonas aeruginosa mutants: implications for susceptibility testing and treatment of chronic infections. Antimicrob Agents Chemother. 2004; 48: 4226-33. [DOI:10.1128/AAC.48.11.4226-4233.2004]
10. Macia MD, Blanquer D, Togores B, Sauleda J, Perez JL, and Oliver A. Hypermutation is a key factor in development of multiple-antimicrobial resistance in Pseudomonas aeruginosa strains causing chronic lung infections. Antimicrob Agents Chemother. 2005; 49: 3382-6. [DOI:10.1128/AAC.49.8.3382-3386.2005]
11. Oliver A, Baquero F, and Blazquez J. The mismatch repair system (mutS, mutL and uvrD genes) in Pseudomonas aeruginosa: molecular characterization of naturally occurring mutants. Mol Microbiol. 2002; 43: 1641-50. [DOI:10.1046/j.1365-2958.2002.02855.x]
12. Ekrami A, and Kalantar E. Bacterial infection in burn patients at a burn hospital in Iran. Indian J Med Res. 2007; 126: 541-4.
13. Estahbanati HK, Kashani PP, Ghanaatpishe F. Frequency of Pseudomonas aeroginosa serotypes in burn wound infections and their resistance to antibiotics. Burns. 2002; 28: 340-8. [DOI:10.1016/S0305-4179(02)00024-4]
14. Kovacs B, Le Gall-David S, Vincent P, Le Bars H, Buffet-Bataillon S, Bonnaure-Mallet M, et al. Is biofilm formation related to the hypermutator phenotype in clinical Enterobacteriaceae isolates? FEMS Microbiol Lett. 2013; 347 (2): 116-22 [DOI:10.1111/1574-6968.12229]
15. Guelfo JR, Rodriguez-Rojas A, Matic I, Blazquez J. A MATE-family efflux pump rescues the Escherichia coli 8-oxoguanine-repair-deficient mutator phenotype and protects against H2O2 killing. PLoS Genetics. 2010; 6 (5). [DOI:10.1371/journal.pgen.1000931]
16. Eschenlauer SC, Coombs GH, Mottram JC. PFPI-like genes are expressed in Leishmania major but are pseudogenes in other Leishmania species. FEMS Microbiol Lett. 2006; 260 (1): 47-54. [DOI:10.1111/j.1574-6968.2006.00303.x]
17. Mandsberg LF, Maciá MD, Bergmann KR, Christiansen LE, Alhede M, Kirkby N, et al. Development of antibiotic resistance and up-regulation of the antimutator gene pfpI in mutator Pseudomonas aeruginosa due to inactivation of two DNA oxidative repair genes (mutY, mutM). FEMS Microbiol Lett. 2011; 324 (1): 28-37. [DOI:10.1111/j.1574-6968.2011.02383.x]
18. Roy A, Kucukural A, and Zhang Y. I-TASSER: a unified platform for automated protein structure and function prediction. Nat Protoc. 2010; 5 (4): 725-38. [DOI:10.1038/nprot.2010.5]
Add your comments about this article : Your username or Email:
CAPTCHA
Send email to the article author

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.