Emerge of NDM-1-Producing Multidrug-Resistant Pseudomo-nas aeruginosa and Co-Harboring of Carbapenemase Genes in South of Iran

  • Ahmad FARAJZADEH SHEIKH 1. Department of Microbiology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran 2. Health Research Institute, Infectious and Tropical Diseases Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran
  • Mojtaba SHAHIN Department of Medical Laboratory Sciences, Faculty of Medical Sciences, Arak Branch, Islamic Azad University, Arak, Iran
  • Leili SHOKOOHIZADEH Department of Microbiology, Faculty of Medicine, Hamadan University of Medical Sciences, Hamedan, Iran
  • Fahimeh GHANBARI Student Research Committee, Shahid Sadoughi University of Medical Sciences, Yazd, Iran
  • Hamid SOLGI Department of Bacteriology, Microbiology Research Center, Pasteur Institute of Iran, Tehran, Iran
  • Fereshteh SHAHCHERAGHI Department of Bacteriology, Microbiology Research Center, Pasteur Institute of Iran, Tehran, Iran
Pseudomonas aeruginosa;, New Delhi metallo-β-lactamase (blaNDM-1);, Modified hodge test (MHT);, Double-disk potentiation tests (DDPT);, Double disk synergy test (DDST)


Background: New Delhi metallo-beta-lactamase-1 (NDM-1) is one of the most important emerging antibiotic resistance. Co-harboring three or four carbapenemases is rare and only a few reports exist in the literature. We described the characteristics of the large epidemic outbreaks and reports co-producing blaNDM-1 with the other carbapenemase genes in P. aeruginosa isolates.

Methods: This present cross-sectional research was conducted on 369 P. aeruginosa isolates obtained from burn and general hospitals within years 2013 to 2016. Beta-lactamase classes A, B and D genes were identified by PCR method. Modified hodge test (MHT), double-disk potentiation tests (DDPT) and double disk synergy test (DDST) were performed for detection carbapenemase and metallo beta-lactamase (MBL) production of blaNDM-1 positive P. aeruginos isolates.

Results: From 236 carbapenem-resistant P. aeruginosa (CRPA), 116 isolates have had MBL genes and twenty-nine isolates were found positive for blaNDM-1. In CRPA isolates, blaIMP-1, blaVIM-2 and blaOXA-10 were identified in 27.5%, 21.1% and 32.2% of isolates respectively, while co-producing blaNDM-1, blaIMP-1, blaOXA-10, co-producing blaNDM-1, blaVIM-2, blaOXA-10 and co-producing blaIMP-1, blaVIM-2 were determined in 11 (4.6%), 8 (3.4%) and 27 (11.4%) of isolates respectively.

Conclusion: The finding of this co-existence of multiple carbapenemase resistance genes is threating for public health. Dipicolinic acid is a superior MBL inhibitor in DDPT antique than EDTA in DDST method for the detection of MBL-blaNDM-1 producing P. aeruginosa.



1. Faghri J, Nouri S, Jalalifar S, Zalipoor M, Halaji M (2018). Investigation of antimicrobial susceptibility, class I and II integrons among Pseudomonas aeruginosa isolates from hospitalized patients in Isfahan, Iran. BMC Res Notes, 11(1):806.
2. Queenan AM, Bush K (2007). Carbapenemases: the versatile beta-lactamases. Clin Microbiol Rev, 20(3):440-58.
3. Jamal WY, Albert MJ, Rotimi VO (2016). High Prevalence of New Delhi Metallo-beta-Lactamase-1 (NDM-1) Producers among Carbapenem-Resistant Enterobacteriaceae in Kuwait. PLoS One, 11(3):e0152638.
4. Farajzadeh Sheikh A, Rostami S, Jolodar A et al (2014). Detection of Metallo-Beta Lactamases Among Carbapenem-Resistant Pseudomonas aeruginosa. Jundishapur J Microbiol, 7(11):e12289.
5. Rezaei A, Fazeli H, Moghadampour M, Halaji M, Faghri J (2018). Determination of antibiotic resistance pattern and prevalence of OXA-type carbapenemases among Acinetobacter baumannii clinical isolates from inpatients in Isfahan, central Iran. Infez Med, 26(1):61-66.
6. Jovcic B, Lepsanovic Z, Begovic J et al (2014). Two copies of blaNDM-1 gene are present in NDM-1 producing Pseudomonas aeruginosa isolates from Serbia. Antonie Van Leeuwenhoek, 105(3):613-8.
7. Paul D, Dhar D, Maurya AP, et al (2016). Occurrence of co-existing bla VIM-2 and bla NDM-1 in clinical isolates of Pseudomonas aeruginosa from India. Ann Clin Microbiol Antimicrob, 15:31.
8. Wang M, Borris L, Aarestrup FM et al (2015). Identification of a Pseudomonas aeruginosa co-producing NDM-1, VIM-5 and VIM-6 metallo-beta-lactamases in Denmark using whole-genome sequencing. Int J Antimicrob Agents, 45(3):324-5.
9. Lavenir R, Jocktane D, Laurent F et al (2007). Improved reliability of Pseudomonas aeruginosa PCR detection by the use of the species-specific ecfX gene target. J Microbiol Methods, 70(1):20-9.
10. Wayne P (2018). Clinical and Laboratory Standards Institute: Performance standards for antimicrobial susceptibility testing: Twenty-fourth informational supplement, M100-S28. Clinical and Laboratory Standards Institute (CLSI), 34(1).
11. Amjad A, Mirza I, Abbasi S et al (2011). Modified Hodge test: A simple and effective test for detection of carbapenemase production. Iran J Microbiol, 3(4):189-93.
12. Qu TT, Zhang JL, Wang J et al (2009). Evaluation of phenotypic tests for detection of metallo-beta-lactamase-producing Pseudomonas aeruginosa strains in China. J Clin Microbiol, 47(4):1136-42.
13. Najar Peerayeh S, Pirhajati Mahabadi R, Pakbaten Toupkanlou S et al (2014). Diversity of beta-lactamases produced by imipenem resistant, Pseudomonas aeruginosa isolates from the bloodstream. Burns, 40(7):1360-4.
14. Yong D, Lee K, Yum JH et al (2002). Imipenem-EDTA disk method for differentiation of metallo-beta-lactamase-producing clinical isolates of Pseudomonas spp. and Acinetobacter spp. J Clin Microbiol, 40(10):3798-801.
15. Yong D, Lee Y, Jeong SH et al (2012). Evaluation of double-disk potentiation and disk potentiation tests using dipicolinic acid for detection of metallo-beta-lactamase-producing pseudomonas spp. and Acinetobacter spp. J Clin Microbiol, 50(10):3227-32.
16. Flateau C, Janvier F, Delacour H et al (2012). Recurrent pyelonephritis due to NDM-1 metallo-beta-lactamase producing Pseudomonas aeruginosa in a patient returning from Serbia, France, 2012. Euro Surveill, 17(45).
17. Van der Bij AK, Pitout JD (2012). The role of international travel in the worldwide spread of multiresistant Enterobacteriaceae. J Antimicrob Chemother, 67(9):2090-100.
18. Jovcic B, Lepsanovic Z, Suljagic V et al (2011). Emergence of NDM-1 metallo-beta-lactamase in Pseudomonas aeruginosa clinical isolates from Serbia. Antimicrob Agents Chemother, 55(8):3929-31.
19. Shokri D, Rabbani Khorasgani M, Fatemi SM et al (2017). Resistotyping, phenotyping and genotyping of New Delhi metallo-beta-lactamase (NDM) among Gram-negative bacilli from Iranian patients. J Med Microbiol, 66(4):402-11.
20. Rizek C, Fu L, Dos Santos LC et al (2014). Characterization of carbapenem-resistant Pseudomonas aeruginosa clinical isolates, carrying multiple genes coding for this antibiotic resistance. Ann Clin Microbiol Antimicrob, 13:43.
21. Farzana R, Shamsuzzaman S, Mamun KZ (2013). Isolation and molecular characterization of New Delhi metallo-beta-lactamase-1 producing superbug in Bangladesh. J Infect Dev Ctries, 7(3):161-8.
22. Malkocoglu G, Aktas E, Bayraktar B et al (2017). VIM-1, VIM-2, and GES-5 Carbapenemases Among Pseudomonas aeruginosa Isolates at a Tertiary Hospital in Istanbul, Turkey. Microb Drug Resist, 23(3):328-34.
23. Johnson AP, Woodford N(2013) . Global spread of antibiotic resistance: the example of New Delhi metallo-β-lactamase (NDM)-mediated carbapenem resistance. J Med Microbiol,62(4):499-513.
24. Mirsalehian A, Feizabadi M, Nakhjavani FA et al (2010). Detection of VEB-1, OXA-10 and PER-1 genotypes in extended-spectrum beta-lactamase-producing Pseudomonas aeruginosa strains isolated from burn patients. Burns, 36(1):70-4.
25. Ranjbar R, Owlia P, Saderi H et al (2011). Characterization of Pseudomonas aeruginosa strains isolated from burned patients hospitalized in a major burn center in Tehran, Iran. Acta Med Iran. 49(10):675-9.
26. Sedighi M, Vaez H, Moghoofeie M, Hadifar S, Oryan G, Faghri J (2015). Molecular detection of metallo-β-lactamase gene blaVIM-1 in imipenem-resistant Pseudomonas aeruginosa strains isolated from hospitalized patients in the hospitals of Isfahan. Adv Biomed Res, 4:57.
27. Cornaglia G, Mazzariol A, Lauretti L et al (2000). Hospital outbreak of carbapenem-resistant Pseudomonas aeruginosa producing VIM-1, a novel transferable metallo-beta-lactamase. Clin Infect Dis, 31(5):1119-25.
28. Yong D, Toleman MA, Giske CG et al (2009). Characterization of a new metallo-β-lactamase gene, blaNDM-1, and a novel erythromycin esterase gene carried on a unique genetic structure in Klebsiella pneumoniae sequence type 14 from India. Antimicrob Agents Chemother, 53(12):5046-54.
29. Kazeminezhad B, Rad AB, Gharib A, Zahedifard S (2017). blaVIM and blaIMP Genes Detection in Isolates of Carbapenem Resistant P. aeruginosa of Hospitalized Patients in Two Hospitals in Iran. Iran J Pathol, 12(4):392-396.
30. Toleman MA, Spencer J, Jones L et al (2012). blaNDM-1 is a chimera likely constructed in Acinetobacter baumannii. Antimicrob Agents Chemother, 56(5):2773-6.
31. Zavascki AP, Barth AL, Gonçalves AL et al (2006).The influence of metallo-β-lactamase production on mortality in nosocomial Pseudomonas aeruginosa infections. J Antimicrob Chemother. 58(2):387-92.
32. Golshani Z, Sharifzadeh A (2013). Prevalence of blaOxa10 Type Beta-lactamase Gene in Carbapenemase Producing Pseudomonas aeruginosa Strains Isolated From Patients in Isfahan. Jundishapur J Microbiol, 6(5).
33. Wei WJ, Yang HF, Ye Y et al (2015). New Delhi Metallo-beta-Lactamase-Mediated Carbapenem Resistance: Origin, Diagnosis, Treatment and Public Health Concern. Chin Med J (Engl), 128(14):1969-76.
34. Shacheraghi F, Shakibaie MR, Noveiri H (2010). Molecular identification of ESBL Genes blaGES-blaVEB-blaCTX-M blaOXA-blaOXA-4, blaOXA-10 andblaPER-in Pseudomonas aeruginosa strains isolated from burn patients by PCR, RFLP and sequencing techniques. Int J Biol life Sci, 3(6):138-42.
35. Poirel L, Walsh TR, Cuvillier V et al (2011). Multiplex PCR for detection of acquired carbapenemase genes. Diagn Microbiol Infect Dis, 70(1):119-23.
36. Nordmann P, Naas T, Poirel L (2011). Global spread of carbapenemase-producing Enterobacteriaceae. Emerg Infect Dis, 17(10):1791-8.
How to Cite
FARAJZADEH SHEIKH A, SHAHIN M, SHOKOOHIZADEH L, GHANBARI F, SOLGI H, SHAHCHERAGHI F. Emerge of NDM-1-Producing Multidrug-Resistant Pseudomo-nas aeruginosa and Co-Harboring of Carbapenemase Genes in South of Iran. Iran J Public Health. 49(5):959-967.
Original Article(s)