Development of multiplex PCR for rapid detection of metallo-β-lactamase genes in clinical isolates of Acinetobacter baumannii
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Abstract
Background and Objectives: Acinetobacter baumannii has been known as a major pathogen causing nosocomial infections. The aim of this study was to develop multiplex PCR for rapid and simultaneous detection of metallo-β-lactamase (MBL) genes in clinical isolates of A. baumannii.
Materials and Methods: In this study, we used three sets of primers to amplify the MBL genes including blaOXA-48, blaOXA-23 and blaNDM. The multiplex PCR assay was optimized for rapid and simultaneous detection of MBL genes in A. baumannii strains recovered from clinical samples.
Results: A. baumannii strains recovered from clinical samples were subjected to the study. The multiplex PCR produced 3 bands of 501 bp for blaOXA-23, 744 bp for blaOXA-48 and 623 bp for blaNDM genes. In addition to, no any cross-reactivity was observed in multiplex PCR.
Conclusion: Based on obtained data, the multiplex PCR had a good specificity without any cross reactivity and it appears that the multiplex PCR is reliable assay for simultaneous detection of MBL genes in A. baumannii strains.
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3. Gordon NC, Wareham DW. Multidrug-resistant Acinetobacter baumannii: mechanisms of virulence and resistance. Int J Antimicrob Agents 2010; 35:219-226.
4. Ranjbar R, Sadeghifard N, Ahmadi A, Izadi M, Zaeimi-Yazdi J, Ghasemi A, et al. Antimicrobial susceptibility and AP-PCR typing of Acinetobacter spp. strains. Iranian J Publ Health 2007; 36: 50-56.
5. Bassetti M, Righi E, Esposito S, Petrosillo N, Nicolini L. Drug treatment for multidrug-resistant Acinetobacter baumannii infections. Future Microbiol 2008; 3:649-660.
6. Mohammadpour B, Rouhi S, Moradi M, Ramazanzadeh R, Saniyi E, Zandi S, et al. Prevalence of metallo-β-lactamases in Acinetobacter baumannii in Iran: A review and meta-analysis. Infect Disord Drug Targets 2019;19:350-361.
7. Cheikh HB, Domingues S, Silveira E, Kadri Y, Rosário N, Mastouri M, et al. Molecular characterization of carbapenemases of clinical Acinetobacter baumannii-calcoaceticus complex isolates from a University Hospital in Tunisia. 3 Biotech 2018; 8:297.
8. Rodríguez CH, Nastro M, Famiglietti A. Carbapenemases in Acinetobacter baumannii. Review of their dissemination in Latin America. Rev Argent Microbiol 2018; 50:327-333.
9. Karah N, Sundsfjord A, Towner K, Samuelsen Ø. Insights into the global molecular epidemiology of carbapenem non-susceptible clones of Acinetobacter baumannii. Drug Resist Updat 2012; 15:237-247.
10. Djahmi N, Dunyach-Remy C, Pantel A, Dekhil M, Sotto A, Lavigne JP. Epidemiology of carbapenemase-producing Enterobacteriaceae and Acinetobacter baumannii in Mediterranean countries. Biomed Res Int 2014; 2014:305784.
11. Safari M, Mozaffari Nejad AS, Bahador A, Jafari R, Alikhani MY. Prevalence of ESBL and MBL encoding genes in Acinetobacter baumannii strains isolated from patients of intensive care units (ICU). Saudi J Biol Sci 2015; 22:424-429.
12. Aksoy MD, Çavuşlu Ş, Tuğrul HM. Investigation of metallo beta lactamases and oxacilinases in carbapenem resistant Acinetobacter baumannii strains isolated from inpatients. Balkan Med J 2015; 32:79-83.
13. Hakemi Vala M, Hallajzadeh M, Hashemi A, Goudarzi H, Tarhani M, Sattarzadeh Tabrizi M, et al. Detection of Ambler class A, B and D ß-lactamases among Pseudomonas aeruginosa and Acinetobacter baumannii clinical isolates from burn patients. Ann Burns Fire Disasters 2014; 27:8-13.
14. Tran DN, Tran HH, Matsui M, Suzuki M, Suzuki S, Shibayama K, et al. Emergence of New Delhi metallo-beta-lactamase 1 and other carbapenemase-producing Acinetobacter calcoaceticus-baumannii complex among patients in hospitals in Ha Noi, Viet Nam. Eur J Clin Microbiol Infect Dis 2017; 36:219-225.
15. Poirel L, Nordmann P. Carbapenem resistance in Acinetobacter baumannii: mechanisms and epidemiology. Clin Microbiol Infect 2006; 12:826-836.
16. Pillonetto M, Arend L, Vespero EC, Pelisson M, Gomes Chagas TP, D'Alincourt Carvalho-Assef AP, et al. First report of NDM-1-producing Acinetobacter baumannii sequence type 25 in Brazil. Antimicrob Agents Chemother 2014; 58: 7592-7594.
17. Nordmann P, Poirel L, Carrër A, Toleman MA, Walsh TR. How to detect NDM-1 producers. J Clin Microbiol 2011; 49:718-721.
18. Shahcheraghi F, Abbasalipour M, Feizabadi M, Ebrahimipour G, Akbari N. Isolation and genetic characterization of metallo-β-lactamase and carbapenamase producing strains of Acinetobacter baumannii from patients at Tehran hospitals. Iran J Microbiol 2011; 3:68-74.
19. Neonakis IK, Spandidos DA, Petinaki E. Confronting multidrug-resistant Acinetobacter baumannii: a review. Int J Antimicrob Agents 2011; 37:102-109.
20. Sadeghifard N, Ranjbar R, Zaeimi J. Antimicrobial susceptibility, plasmid profiles, and RAPD-PCR typing of Acinetobacter bacteria. Asian Biomed 2010;4:901-911.
21. Zavascki AP, Carvalhaes CG, Picão RC, Gales AC. Multidrug-resistant Pseudomonas aeruginosa and Acinetobacter baumannii: resistance mechanisms and implications for therapy. Expert Rev Anti Infect Ther 2010; 8:71-93.
22. Hu Q, Hu Z, Li J, Tian B, Xu H, Li J. Detection of OXA-type carbapenemases and integrons among carbapenem-resistant Acinetobactor baumannii in a teaching hospital in China. J Basic Microbiol 2011; 51:467-472.
23. Poirel L, Walsh TR, Cuvillier V, Nordmann P. Multiplex PCR for detection of acquired carbapenemase genes. Diagn Microbiol Infect Dis 2011;70:119-123.
24. Aksoy MD, Çavuşlu Ş, Tuğrul HM. Investigation of metallo beta lactamases and oxacilinases in carbapenem resistant Acinetobacter baumannii strains isolated from inpatients. Balkan Med J 2015; 32:79-83.
25. Mostachio AK, van der Heidjen I, Rossi F, Levin AS, Costa SF. Multiplex PCR for rapid detection of genes encoding oxacillinases and metallo-beta-lactamases in carbapenem-resistant Acinetobacter spp. J Med Microbiol 2009; 58:1522-1524.
26. Woodford N, Turton JF, Livermore DM. Multiresistant Gram-negative bacteria: the role of high-risk clones in the dissemination of antibiotic resistance. FEMS Microbiol Rev 2011;35:736-755.
27. Mlynarcik P, Roderova M, Kolar M. Primer evaluation for PCR and its application for detection of carbapenemases in Enterobacteriaceae. Jundishapur J Microbiol 2016; 9(1): e29314.
28. Kamolvit W, Higgins PG, Paterson DL, Seifert H. Multiplex PCR to detect the genes encoding naturally occurring oxacillinases in Acinetobacter spp. J Antimicrob Chemother 2014;69:959-963.
2. Nowak P, Paluchowska P. Acinetobacter baumannii: biology and drug resistance - role of carbapenemases. Folia Histochem Cytobiol 2016; 54:61-74.
3. Gordon NC, Wareham DW. Multidrug-resistant Acinetobacter baumannii: mechanisms of virulence and resistance. Int J Antimicrob Agents 2010; 35:219-226.
4. Ranjbar R, Sadeghifard N, Ahmadi A, Izadi M, Zaeimi-Yazdi J, Ghasemi A, et al. Antimicrobial susceptibility and AP-PCR typing of Acinetobacter spp. strains. Iranian J Publ Health 2007; 36: 50-56.
5. Bassetti M, Righi E, Esposito S, Petrosillo N, Nicolini L. Drug treatment for multidrug-resistant Acinetobacter baumannii infections. Future Microbiol 2008; 3:649-660.
6. Mohammadpour B, Rouhi S, Moradi M, Ramazanzadeh R, Saniyi E, Zandi S, et al. Prevalence of metallo-β-lactamases in Acinetobacter baumannii in Iran: A review and meta-analysis. Infect Disord Drug Targets 2019;19:350-361.
7. Cheikh HB, Domingues S, Silveira E, Kadri Y, Rosário N, Mastouri M, et al. Molecular characterization of carbapenemases of clinical Acinetobacter baumannii-calcoaceticus complex isolates from a University Hospital in Tunisia. 3 Biotech 2018; 8:297.
8. Rodríguez CH, Nastro M, Famiglietti A. Carbapenemases in Acinetobacter baumannii. Review of their dissemination in Latin America. Rev Argent Microbiol 2018; 50:327-333.
9. Karah N, Sundsfjord A, Towner K, Samuelsen Ø. Insights into the global molecular epidemiology of carbapenem non-susceptible clones of Acinetobacter baumannii. Drug Resist Updat 2012; 15:237-247.
10. Djahmi N, Dunyach-Remy C, Pantel A, Dekhil M, Sotto A, Lavigne JP. Epidemiology of carbapenemase-producing Enterobacteriaceae and Acinetobacter baumannii in Mediterranean countries. Biomed Res Int 2014; 2014:305784.
11. Safari M, Mozaffari Nejad AS, Bahador A, Jafari R, Alikhani MY. Prevalence of ESBL and MBL encoding genes in Acinetobacter baumannii strains isolated from patients of intensive care units (ICU). Saudi J Biol Sci 2015; 22:424-429.
12. Aksoy MD, Çavuşlu Ş, Tuğrul HM. Investigation of metallo beta lactamases and oxacilinases in carbapenem resistant Acinetobacter baumannii strains isolated from inpatients. Balkan Med J 2015; 32:79-83.
13. Hakemi Vala M, Hallajzadeh M, Hashemi A, Goudarzi H, Tarhani M, Sattarzadeh Tabrizi M, et al. Detection of Ambler class A, B and D ß-lactamases among Pseudomonas aeruginosa and Acinetobacter baumannii clinical isolates from burn patients. Ann Burns Fire Disasters 2014; 27:8-13.
14. Tran DN, Tran HH, Matsui M, Suzuki M, Suzuki S, Shibayama K, et al. Emergence of New Delhi metallo-beta-lactamase 1 and other carbapenemase-producing Acinetobacter calcoaceticus-baumannii complex among patients in hospitals in Ha Noi, Viet Nam. Eur J Clin Microbiol Infect Dis 2017; 36:219-225.
15. Poirel L, Nordmann P. Carbapenem resistance in Acinetobacter baumannii: mechanisms and epidemiology. Clin Microbiol Infect 2006; 12:826-836.
16. Pillonetto M, Arend L, Vespero EC, Pelisson M, Gomes Chagas TP, D'Alincourt Carvalho-Assef AP, et al. First report of NDM-1-producing Acinetobacter baumannii sequence type 25 in Brazil. Antimicrob Agents Chemother 2014; 58: 7592-7594.
17. Nordmann P, Poirel L, Carrër A, Toleman MA, Walsh TR. How to detect NDM-1 producers. J Clin Microbiol 2011; 49:718-721.
18. Shahcheraghi F, Abbasalipour M, Feizabadi M, Ebrahimipour G, Akbari N. Isolation and genetic characterization of metallo-β-lactamase and carbapenamase producing strains of Acinetobacter baumannii from patients at Tehran hospitals. Iran J Microbiol 2011; 3:68-74.
19. Neonakis IK, Spandidos DA, Petinaki E. Confronting multidrug-resistant Acinetobacter baumannii: a review. Int J Antimicrob Agents 2011; 37:102-109.
20. Sadeghifard N, Ranjbar R, Zaeimi J. Antimicrobial susceptibility, plasmid profiles, and RAPD-PCR typing of Acinetobacter bacteria. Asian Biomed 2010;4:901-911.
21. Zavascki AP, Carvalhaes CG, Picão RC, Gales AC. Multidrug-resistant Pseudomonas aeruginosa and Acinetobacter baumannii: resistance mechanisms and implications for therapy. Expert Rev Anti Infect Ther 2010; 8:71-93.
22. Hu Q, Hu Z, Li J, Tian B, Xu H, Li J. Detection of OXA-type carbapenemases and integrons among carbapenem-resistant Acinetobactor baumannii in a teaching hospital in China. J Basic Microbiol 2011; 51:467-472.
23. Poirel L, Walsh TR, Cuvillier V, Nordmann P. Multiplex PCR for detection of acquired carbapenemase genes. Diagn Microbiol Infect Dis 2011;70:119-123.
24. Aksoy MD, Çavuşlu Ş, Tuğrul HM. Investigation of metallo beta lactamases and oxacilinases in carbapenem resistant Acinetobacter baumannii strains isolated from inpatients. Balkan Med J 2015; 32:79-83.
25. Mostachio AK, van der Heidjen I, Rossi F, Levin AS, Costa SF. Multiplex PCR for rapid detection of genes encoding oxacillinases and metallo-beta-lactamases in carbapenem-resistant Acinetobacter spp. J Med Microbiol 2009; 58:1522-1524.
26. Woodford N, Turton JF, Livermore DM. Multiresistant Gram-negative bacteria: the role of high-risk clones in the dissemination of antibiotic resistance. FEMS Microbiol Rev 2011;35:736-755.
27. Mlynarcik P, Roderova M, Kolar M. Primer evaluation for PCR and its application for detection of carbapenemases in Enterobacteriaceae. Jundishapur J Microbiol 2016; 9(1): e29314.
28. Kamolvit W, Higgins PG, Paterson DL, Seifert H. Multiplex PCR to detect the genes encoding naturally occurring oxacillinases in Acinetobacter spp. J Antimicrob Chemother 2014;69:959-963.
| Files | ||
| Issue | Vol 12 No 2 (2020) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v12i2.2615 | |
| Keywords | ||
| Acinetobacter baumannii; Metallo-β-lactamase; Multiplex polymerase chain reaction | ||
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This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
How to Cite
1.
Ranjbar R, Zayeri S, Mirzaie A. Development of multiplex PCR for rapid detection of metallo-β-lactamase genes in clinical isolates of Acinetobacter baumannii. Iran J Microbiol. 2020;12(2):107-112.
