Antibiotyping and Genotyping of Pseudomonas aeruginosa Strains Isolated from Mottahari Hospital in Tehran, Iran by ERIC-PCR

Authors
H. Mohammadi Barzelighi 1
H. Fazeli 1
B. Nasr Esfahani 1
M. Sattarzadeh 2
H. Mohammadi Barzelighi 1
1 Department of Bacteriology, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, IR Iran
2 Motahhari Burn Hospital, Iran University of Medical Sciences, Tehran, IR Iran
Abstract
Background: Pseudomonas aeruginosa has become the most common cause of infections in burn patients. The aim of this study was to investigate the antibiotyping and genotyping of P. aeruginosa strains isolated from burn patients in Mottahari hospital during June-October 2016.
Materials and Methods: A total of 78 P. aeruginosa strains were collected from wound infected patients. Identification of the isolates was performed by biochemical tests and confirmed by specific 16srDNA PCR. Antimicrobial susceptibility testing was done by disk diffusion method according to the CLSI guidelines. The isolates were then evaluated for genotyping by ERIC-PCR.
Results: From a total of 78 collected isolates, 77 isolates (98.7%) were confirmed as P. aeruginosa by specific PCR. We found 4 antibiotypes. The highest resistance was observed to imipenem and gentamicin (~100%), and the most sensitivity was shown to colistin (100%). Overall, MDR phenotype was observed in most of the isolates (98.7%). The PCR of ERIC box produced 52 different patterns and 3 main clusters. Also, 59 (83%), 2 (3%), and 9 (13%) isolates were included in Cluster A, B, and C, respectively, and Cluster A was the predominant ERIC profile.
Conclusion: The high resistance to antibiotics in our study may be due to their abundant use as the prophylactic or treatment regimen in wound infections. So appropriate use of antibiotics seems necessary, and colistin is a proper choice for treatment of burn infection. In genotyping, 3 main clusters and 52 different patterns were shown. A majority of the P. aeruginosa strains isolated from burn patients were related and belonged to Cluster A.

Keywords

1.   Rossolini G, Mantengoli E. Treatment and control of severe infections caused by multiresistant Pseudomonas aeruginosa. Clin Microbiol Infect. 2005; 11(Suppl 4):17-32.
2.   Lee J-Y, Ko KS. Mutations and expression of PmrAB and PhoPQ related with colistin resistance in Pseudomonas aeruginosa clinical isolates. Diagno Microbiol Infect Dis. 2014; 78(3):271-6.
3.   Patzer J, Toleman MA, Deshpande LM, Kamińska W, Dzierżanowska D, Bennett PM, et al. Pseudomonas aeruginosa strains harbouring an unusual blaVIM-4 gene cassette isolated from hospitalized children in Poland (1998–2001). J Antimicrob Chemother. 2004; 53(3):451-6.
4.   Shahcheraghi F, Badmasti F, Feizabadi MM. Molecular characterization of Class 1 integrons in MDR Pseudomonas aeruginosa isolated from clinical settings in Iran, Tehran. FEMS Immunol Med Microbiol. 2010; 58(3):421-5.
5.   Gales AC, Castanheira M, Jones RN, Sader HS. Antimicrobial resistance among gram-negative bacilli isolated from Latin America: results from SENTRY Antimicrobial Surveillance Program (Latin America, 2008–2010). Diagn Microbiol Infect Dis. 2012; 73(4):354-60.
6.   Xu Z, Li L, Shirtliff M, Peters B, Li B, Peng Y, et al. Resistance Class 1 integron in clinical methicillin‐resistant Staphylococcus aureus strains in southern China, 2001–2006. Clin Microbiol Infect. 2011; 17(5):714-8.
7.   Valadbeigi H, Rafiei Tabatabaei R, Malek A, Sekawi Z, Raftari M, Parvaneh K, et al. Genomic diversity and virulence genes among clinical isolates of Pseudomonas aeruginosa. Clin Lab. 2014; 60:363-7
8.   Syrmis MW, O'carroll MR, Sloots TP, Coulter C, Wainwright CE, Bell SC, et al. Rapid genotyping of Pseudomonas aeruginosa isolates harboured by adult and paediatric patients with cystic fibrosis using repetitive-element-based PCR assays. J Med Microbiol. 2004; 53(Pt 11):1089-96.
9.   Nanvazadeh F, Khosravi AD, Zolfaghari MR, Parhizgari N. Genotyping of Pseudomonas aeruginosa strains isolated from burn patients by RAPD-PCR. Burns. 2013; 39(7):1409-13.
10. Spilker T, Coenye T, Vandamme P, LiPuma JJ. PCR-based assay for differentiation of Pseudomonas aeruginosa from other Pseudomonas species recovered from cystic fibrosis patients. J Clin Microbiol. 2004; 42(5):2074-9.
11. Wayne PA. Clinical and laboratory standards institute; 2007. Performance standards for antimicrobial susceptibility testing. CLSI document M100-S17.
12. Vitkauskiene A, Skrodeniene E, Dambrauskiene A, Macas A, Sakalauskas R. Pseudomonas aeruginosa bacteremia: resistance to antibiotics, risk factors, and patient mortality. Medicina (Kaunas). 2010; 46(7):490-5.
13. Falagas ME, Koletsi PK, Bliziotis IA. The diversity of definitions of multidrug-resistant (MDR) and pandrug-resistant (PDR) Acinetobacter baumannii and Pseudomonas aeruginosa. J Med Microbiol. 2006; 55(Pt 12):1619-29.
14. Mansury D, Motamedifar M, Sarvari J, Shirazi B, Khaledi A. Antibiotic susceptibility pattern and identification of extended spectrum β-lactamases (ESBLs) in clinical isolates of Klebsiella pneumoniae from Shiraz, Iran. Iran J Microbiol. 2016; 8(1):55.
15. Ranjbar R, Owlia P, Saderi H, Mansouri S, Jonaidi-Jafari N, Izadi M, et al. Characterization of Pseudomonas aeruginosa strains isolated from burned patients hospitalized in a major burn center in Tehran, Iran. Acta Med Iran. 2011;49(10):675-9.
16. Rajaie S, Mohammadi Sichani M, Mohammad Yousefi H. Study on the inhibitory activity of zinc oxide nanoparticles against Pseudomonas aeruginosa isolated from burn wounds. Qom Univ Med Sci J. 2015; 9(1):30-7.
17. Fazeli H, Havaei SA, Solgi H, Shokri D, Motallebirad T. Pattern of antibiotic resistance in Pesudomonas aeruginosa isolated from intensive care unit, Isfahan, Iran. J Isfahan Med Sch. 2013; 31(232):433-438.
18. Şener B, Köseoğlu Ö, Özçelik U, Günalp A. Epidemiology of chronic Pseudomonas aeruginosa infections in cystic fibrosis. Int J Med Microbiol. 2001; 291(5):387-93.
19. Wolska K, Szweda P. A comparative evaluation of PCR ribotyping and ERIC PCR for determining the diversity of clinical Pseudomonas aeruginosa isolates. Pol J Microbiol. 2008; 57(2):157-63.
20. Khosravi AD, Hoveizavi H, Mohammadian A, Farahani A, Jenabi A. Genotyping of multidrug-resistant strains of Pseudomonas aeruginosa isolated from burn and wound infections by ERIC-PCR. Acta Cir Bras.2016; 31(3):206-11.
21. Ghorbanalizadgan M, Bakhshi B, Lili AK, Najar-Peerayeh S, Nikmanesh B. A molecular survey of Campylobacter jejuni and Campylobacter coli virulence and diversity. Iran Biomed J. 2014;18(3):158-164.