Volume 1, Issue 2 (Summer-Fall 2018)                   Mod Med Lab J 2018, 1(2): 68-76 | Back to browse issues page

XML Print

Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
Ramin B, Asadpour L, Forouhesh Tehrani H, Amirmozafari N. Detection and Distribution of various HLAR Gene in Enterococcus faecalis and Enterococcus faecium by Multiplex-PCR. Mod Med Lab J 2018; 1 (2) :68-76
URL: http://modernmedlab.com/article-1-50-en.html
Detection and Distribution of various HLAR Gene in Enterococcus faecalis and Enterococcus faecium by Multiplex-PCR
Bahar Ramin , Leyla Asadpour , Homa Forouhesh Tehrani , Nour Amirmozafari
Abstract:   (5508 Views)

Background: In recent years, three new aminoglycoside resistance genes such as aph (3ʹ)-IIIa and ant (4ʹ)-Ia, that encode for the APH (3ʹ) and ANT (4ʹ) have also been identified. The aim of this study was to come up with a multiplex-PCR procedure for detection of aac(6ʹ)-Ie–aph(2ʹʹ)-Ia, aph(3ʹ)-IIIa, ant(4ʹ)-Ia genes in the Enterococcus spp. clinical isolates.

Material and Method: 100 samples were isolated from various specimens, from various hospitals in Tehran, Iran. The grown colonies were identified by standard biochemical and disc diffusion tests. Multiplex-PCR for aac (6’)-Ie -aph(2’’)-Ia , aph(3ʹ)-IIIa, ant(4ʹ)-Ia genes amplification were performed in order to confirm bacterial colonies as Enterococcus spp.

Results: Eighty four (84%) Enterococcus spp. isolates were collected from the 100 specimens. The highest and lowest isolates were related to urine (48%) and sputum (2%). Antibiotic susceptibility test results showed that the highest and lowest resistance was related to tetracycline and nitrofurantoin, respectively. Multiplex PCR results revealed that aac (6ʹ)-Ie-aph (2ʹʹ)-Ia, ant (4ʹ)-Ia and aph (3ʹ)-IIIa genes were present in 6% of the isolated bacteria from the urine, 2% from the wound and 1% from the pleural samples. the aac (6ʹ)-Ie-aph (2ʹʹ)-Ia and aph (3ʹ)-IIIa genes were present in 25% of the isolated strains from the urine, 3% from the wound and 2% from the plural specimens. Nine percent of the strains were isolated from the urine, 3% from the wound and 1% from the plural were positive for aac (6ʹ)-Ie-aph (2ʹʹ)-Ia and ant (4ʹ)-Ia genes.

Conclusion: we had observed enterococci isolates with phenotypic resistance to HLAR and demonstrated aac(6′)-Ie-aph(2′′)-Ia and aph(3′)-IIIa genes more frequently occurring than other genes. A collection of AMEs are accountable for HLAR status among Enterococcus species. The aac (6ʹ)-Ie-aph (2ʹʹ)-Ia gene was detected more frequently than the other genes.

Keywords: Enterococcus spp., Antimicrobial Drug Resistance, Multiplex-PCR
Full-Text [PDF 675 kb]   (3087 Downloads)    
Type of Study: Original Research Article | Subject: Clinical Microbiology
References
1. Padmasini E, Padmaraj R, Ramesh SS. High level aminoglycoside resistance and distribution of aminoglycoside resistant genes among clinical isolates of Enterococcus species in Chennai, India. ScientificWorldJournal. 2014;2014:329157. [DOI:10.1155/2014/329157] [PMID] [PMCID]
2. Karlowsky JA, Jones ME, Draghi DC, Thornsberry C, Sahm DF, Volturo GA. Prevalence and antimicrobial susceptibilities of bacteria isolated from blood cultures of hospitalized patients in the United States in 2002. Ann Clin Microbiol Antimicrob. 2004;3:7. [DOI:10.1186/1476-0711-3-7] [PMID] [PMCID]
3. Kobayashi N, Alam M, Nishimoto Y, Urasawa S, Uehara N, Watanabe N. Distribution of aminoglycoside resistance genes in recent clinical isolates of Enterococcus faecalis, Enterococcus faecium and Enterococcus avium. Epidemiol Infect. 2001;126(2):197-204. [DOI:10.1017/S0950268801005271] [PMID] [PMCID]
4. Hasani A, Sharifi Y, Ghotaslou R, Naghili B, Hasani A, Aghazadeh M, et al. Molecular screening of virulence genes in high-level gentamicin-resistant Enterococcus faecalis and Enterococcus faecium isolated from clinical specimens in Northwest Iran. Indian J Med Microbiol. 2012;30(2):175-81. [DOI:10.4103/0255-0857.96687] [PMID]
5. Shinde RS, Koppikar GV, Oommen S. Characterization and antimicrobial susceptibility pattern of clinical isolates of Enterococci at a tertiary care hospital in Mumbai, India. Annals of Tropical Medicine and Public Health. 2012;5(2):85. [DOI:10.4103/1755-6783.95956]
6. Sharifi Y, Hasani A, Ghotaslou R, Naghili B, Aghazadeh M, Milani M, et al. Virulence and antimicrobial resistance in enterococci isolated from urinary tract infections. Adv Pharm Bull. 2013;3(1):197-201. [PMID] [PMCID]
7. Sakoulas G, Nonejuie P, Nizet V, Pogliano J, Crum-Cianflone N, Haddad F. Treatment of high-level gentamicin-resistant Enterococcus faecalis endocarditis with daptomycin plus ceftaroline. Antimicrob Agents Chemother. 2013;57(8):4042-5. [DOI:10.1128/AAC.02481-12] [PMID] [PMCID]
8. Shaw KJ, Rather PN, Hare RS, Miller GH. Molecular genetics of aminoglycoside resistance genes and familial relationships of the aminoglycoside-modifying enzymes. Microbiol Rev. 1993;57(1):138-63. [PMID] [PMCID]
9. Vakulenko SB, Donabedian SM, Voskresenskiy AM, Zervos MJ, Lerner SA, Chow JW. Multiplex PCR for detection of aminoglycoside resistance genes in enterococci. Antimicrob Agents Chemother. 2003;47(4):1423-6. [DOI:10.1128/AAC.47.4.1423-1426.2003] [PMID] [PMCID]
10. Kadam P, Bhalerao S. Sample size calculation. Int J Ayurveda Res. 2010;1(1):55-7. [DOI:10.4103/0974-7788.59946] [PMID] [PMCID]
11. Emaneini M, Aligholi M, Aminshahi M. Characterization of glycopeptides, aminoglycosides and macrolide resistance among Enterococcus faecalis and Enterococcus faecium isolates from hospitals in Tehran. Pol J Microbiol. 2008;57(2):173-8. [PMID]
12. Jamet E, Akary E, Poisson MA, Chamba JF, Bertrand X, Serror P. Prevalence and characterization of antibiotic resistant Enterococcus faecalis in French cheeses. Food Microbiol. 2012;31(2):191-8. [DOI:10.1016/j.fm.2012.03.009] [PMID]
13. Conde-Estevez D, Grau S, Albanell J, Terradas R, Salvado M, Knobel H. Clinical characteristics and outcomes of patients with vancomycin-susceptible Enterococcus faecalis and Enterococcus faecium bacteraemia in cancer patients. Eur J Clin Microbiol Infect Dis. 2011;30(1):103-8. [DOI:10.1007/s10096-010-1029-5] [PMID]
14. Gordon S, Swenson JM, Hill BC, Pigott NE, Facklam RR, Cooksey RC, et al. Antimicrobial susceptibility patterns of common and unusual species of enterococci causing infections in the United States. Enterococcal Study Group. J Clin Microbiol. 1992;30(9):2373-8. [PMID] [PMCID]
15. Olawale KO, Fadiora SO, Taiwo SS. Prevalence of hospital-acquired enterococci infections in two primary-care hospitals in osogbo, southwestern Nigeria. Afr J Infect Dis. 2011;5(2):40-6. [DOI:10.4314/ajid.v5i2.66513]
16. Liu Y, Cao B, Gu L, Wang H. Molecular characterization of vancomycin-resistant enterococci in a Chinese hospital between 2003 and 2009. Microb Drug Resist. 2011;17(3):449-55. [DOI:10.1089/mdr.2011.0025] [PMID]
17. Chow JW. Aminoglycoside resistance in enterococci. Clinical Infectious Diseases. 2000;31(2):586-9. [DOI:10.1086/313949] [PMID]
18. Leclercq R, Dutka-Malen S, Brisson-Noel A, Molinas C, Derlot E, Arthur M, et al. Resistance of enterococci to aminoglycosides and glycopeptides. Clin Infect Dis. 1992;15(3):495-501. [DOI:10.1093/clind/15.3.495] [PMID]
19. Lall N, Basak S. HIGH LEVEL AMINOGLYCOSIDE RESISTANT ENTEROCOCCUS SPECIES: A STUDY. International Journal of Current Research and Review. 2014;6(3):16-21.
20. Hollenbeck BL, Rice LB. Intrinsic and acquired resistance mechanisms in enterococcus. Virulence. 2012;3(5):421-33. [DOI:10.4161/viru.21282] [PMID] [PMCID]
21. Vignaroli C, Zandri G, Aquilanti L, Pasquaroli S, Biavasco F. Multidrug-resistant enterococci in animal meat and faeces and co-transfer of resistance from an Enterococcus durans to a human Enterococcus faecium. Curr Microbiol. 2011;62(5):1438-47. [DOI:10.1007/s00284-011-9880-x] [PMID]
22. Behnood A, Farajnia S, Moaddab SR, Ahdi-Khosroshahi S, Katayounzadeh A. Prevalence of aac(6')-Ie-aph(2'')-Ia resistance gene and its linkage to Tn5281 in Enterococcus faecalis and Enterococcus faecium isolates from Tabriz hospitals. Iran J Microbiol. 2013;5(3):203-8. [PMID] [PMCID]
23. Feizabadi MM, Shokrzadeh L, Sayady S, Asadi S. Transposon Tn5281 is the main distributor of the aminoglycoside modifying enzyme gene among isolates of Enterococcus faecalis in Tehran hospitals. Can J Microbiol. 2008;54(10):887-90. [DOI:10.1139/W08-073] [PMID]
24. Zarrilli R, Tripodi MF, Di Popolo A, Fortunato R, Bagattini M, Crispino M, et al. Molecular epidemiology of high-level aminoglycoside-resistant enterococci isolated from patients in a university hospital in southern Italy. J Antimicrob Chemother. 2005;56(5):827-35. [DOI:10.1093/jac/dki347] [PMID]
25. Vigani AG, Oliveira AM, Bratfich OJ, Stucchi RS, Moretti ML. Clinical, epidemiological, and microbiological characteristics of bacteremia caused by high-level gentamicin-resistant Enterococcus faecalis. Braz J Med Biol Res. 2008;41(10):890-5. [DOI:10.1590/S0100-879X2008001000010] [PMID]
26. Keddy KH, Klugman KP, Liebowitz LD. Incidence of high-level gentamicin resistance in enterococci at Johannesburg Hospital. S Afr Med J. 1996;86(10):1273-6. [PMID]
27. Kuzucu C, Cizmeci Z, Durmaz R, Durmaz B, Ozerol IH. The prevalence of fecal colonization of enterococci, the resistance of the isolates to ampicillin, vancomycin, and high-level aminoglycosides, and the clonal relationship among isolates. Microb Drug Resist. 2005;11(2):159-64. [DOI:10.1089/mdr.2005.11.159] [PMID]
28. Sahm DF, Boonlayangoor S, Schulz JE. Detection of high-level aminoglycoside resistance in enterococci other than Enterococcus faecalis. J Clin Microbiol. 1991;29(11):2595-8. [PMID] [PMCID]
Add your comments about this article : Your username or Email:
CAPTCHA
Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

© 2024 CC BY-NC 4.0 | Modern Medical Laboratory Journal