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Determination of clonal relationships and virulence genes of vancomycin-resistant enterococcus spp. isolated from colonized and infected patients

Year 2019, Volume: 44 Issue: 4, 1442 - 1449, 29.12.2019
https://doi.org/10.17826/cumj.552522

Abstract

Purpose: In this study, we investigated the possible relationship between virulence factors and clonal relationship between Vancomycin-resistant colonization and infection isolates. 

Materials and Methods: A total of 156 Vancomycin-resistant Enterococcus spp. (VRE) were collected and grouped as infection and colonization isolates. A multiplex polymerase chain reaction analysis was performed to screen specific virulence genes (esp, hyl, asa1, cylA and gelE) and vancomycin resistance genes (vanA, vanB). The clonal relationship among isolates was investigated by the Pulsed Field Gel Electrophoresis (PFGE) method. 

Results: The vanA gene was determined in 86 infection and 64 colonization isolates. esp was the most common virulence gene for both groups (55.8% and 56.25%), followed by hyl (51.1% and 35.9%) that statistically differed between the two groups. The 150 E. faecium isolates carrying the vanA resistance gene were divided into 24 main clusters (A-Y) in PFGE analysis. 

Conclusion: When the distribution of virulence genes or genes combinations of the infection and colonization groups was examined, there was no statistical significance of cluster distribution in the two groups. However, hyl gene was found to be more common in infection isolates.


References

  • Eliopoulos GM, Gold HS. Vancomycin-Resistant Enterococci: Mechanisms and Clinical Observations. Clin Infect Dis. 2001; 33: 210-219.
  • Sujatha S, Praharaj I. Glycopeptide Resistance in Gram-Positive Cocci: A Review. Interdis Persp Infect Dis. 2012; 781679: 10.
  • Mundy LM, Sahm DF, Gilmore. Relationships between Enterococcal Virulence and Antimicrobial Resistance. Clin Microbiol Rev. 2000; 13: 513–522.
  • Çetinkaya Y, Falk P, Mayhall CG. Vancomycin-Resistant Enterococi. Clin Microbiol Rev. 2000; 13: 686-707.
  • Leclercg R, Courvalin P. Resistance to Glycopeptides in Enterococci. Clin. Infect Dis. 1997; 24: 545-56.
  • Van der Heijden KM, Van der Heijden IM, Galvao FH, Lopes CG, Costa SF, Abdala E, D’Albuquerque LA, Levin AS. Intestinal Translocation of Clinical Isolates of Vancomycin Resistant Enterococcus faecalis and ESBL-Producing Escherichia coli in a Rat Model of Bacterial Colonization and Liver Ischemia/Reperfusion Injury. PLOS One. 2014; 9: e108453.
  • Higuita NIA, Huycke MM. Enterococcal Disease, Epidemiology, and Implications for Treatment. In: Gilmore MS, Clewell DB, Ike Y, et al. editors. Enterococci: From Commensals to Leading Causes of Drug Resistant Infection. Boston: Massachusetts Eye and Ear Infirmary. 2014 Available from: http://www.ncbi.nlm.nih.gov/books/NBK190429/.
  • Terkuran M, Erginkaya Z, Ünal E, Güran M, Kızılyıldırım S, Ugur G, Köksal F. The Relationship between Virulence Factors and Vancomycin Resistance among Enterococci collected from food and human samples in Southern Turkey. Ankara Üniv Vet Fak Derg. 2014; 61: 133-140.
  • Togay OS, Keskin AÇ, Açık L, Temiz A. Virulence Genes, Antibiotic Resistance and Plasmid Profiles of Enterococcus faecalis and Enterococcus faecium from Naturally Fermented Turkish Foods J Appl. Microbiol. 2010; 109: 1084-1092
  • Garcia-Migura L, Pleydell E, Barnes S, Davies RH, Liebana E. Characterization of Vancomycin-Resistant Enterococcus faecium Isolates from Broiler Poultry and Pig Farms in England and Wales. J Clin Microbiol 2005; 43: 3283–3289.
  • Karakeçe E, Çiftçi IH., Aşık G. Vankomisin Dirençli Enterokoklarda Direncin Moleküler Yöntemlerle Araştırılması. Ankem Derg. 2013; 27: 135-139.
  • Corso AC, Gagetti PS, Rodrı´Guez MM, Melano RG, Ceriana PR, Faccone DF, Galas M.F. Molecular Epidemiology of Vancomycin-Resistant Enterococcus faecium in Argentina. Inter J Inf Dis. 2007; 11: 69-75.
  • Mirzaei B, Farivar TN, Juhari P, Mehr MA, Babaei R. Investigation of the Prevalence of vanA and vanB Genes in Vancomycin Resistant Enterococcus (VRE) by TaqMan RealTime PCR Assay. J Mic Inf Dis. 2013; 3: 192-198.
  • Domingo MC, Huletsky A, Giroux R, Boissinot K, Picard FJ, Lebel P, Ferraro MJ, Bergeron MG. High Prevalence of Glycopeptide Resistance Genes vanB, vanD, and vanG not Associated With Enterococci in Human Fecal Flora. Antimicrob Agents Ch. 2005; 49: 4784–4786.
  • Rice LB, Lakticová V, Carias LL, Rudin S, Hutton R, Marshall SH. Transferable Capacity for Gastrointestinal Colonization in Enterococcus faecium in a Mouse Model. J Infect Dis. 2009; 199(3): 342-349.
  • Leendertse M, Willems RJL, Oei GA, Florquin S, Bonten MJM., Van der Poll M. Intestinal Enterococcus faecium colonization Improves Host Defense During Polymicrobial Peritonitis. J Infect Dis. 2009; 200: 735-744.
  • Diani M, Esiyok OG, Ariafar MN, Yuksel FN, Altuntas EG, Akcelik F. The Interactions between esp, fsr, gelE genes and Biofilm Formation and PFGE analysis of Clinical Enterococcus faecium strains. African J Microbiol Res. 2014; 8: 129-137.
  • Camargo ILBC, Gilmore MS, Darini ALC. Multilocus sequence typing and analysis of putative virulence factors in vancomycin-resistant and vancomycin-sensitive Enterococcus faecium strains isolated in Brazil. Clin Microbiol Infect. 2006; 11: 1130.
  • Çopur ŞS, Şahin F, Göçmen JS. Determination of Virulence and Multidrug Resistance Genes with Polymerase Chain Reaction Method in Vancomycin Sensitive and Resistant Enterococci Isolated from Clinical Samples. Turk J Med Sci. 2016; 46(3): 877-91.
  • Freitas AR, Tedim AP, Novais C, Ruiz-Garbajosa P, Werner G, Laverde-Gomez JA, Cantón R, Peixe L, Baquero F, Coque TM. Global Spread of the hyl(Efm) Colonization-Virulence Gene in Megaplasmids of the Enterococcus faecium CC17 polyclonal subcluster. Antimicrob Agents Chemother. 2010; 54(6): 2660-2665.
  • Vankerckhoven V, Van Autgaerden T, Vael C, Lammens C, Chapelle S, Rossi R, Jabes D, Goossens H. Development of a Multiplex PCR for the Detection of asa1, gelE, cylA, esp, and hyl genes in Enterococci and Survey for Virulence Determinants among European Hospital Isolates of Enterococcus faecium. J Clin Microbiol. 2004; 42: 4473-4479.

Kolonize ve infekte hastalardan izole edilen vankomisin dirençli enterococcus spp.’nin virülans genleri ve klonal ilişkilerinin belirlenmesi

Year 2019, Volume: 44 Issue: 4, 1442 - 1449, 29.12.2019
https://doi.org/10.17826/cumj.552522

Abstract

Amaç: Bu çalışmada, Vankomisine dirençli kolonizasyon ve enfeksiyon izolatları arasındaki klonal ilişki  ve virülans faktörleri araştırılmıştır.

Gereç ve Yöntem: Toplam 156 Vankomisine dirençli Enterococcus spp. (VRE) izole edildi ve enfeksiyon ve kolonizasyon izolatları olarak gruplandı. Spesifik virülans genlerini (esp, hyl, asa1, cylA ve gelE) ve vankomisin direnç genlerini (vanA, vanB) tespit etmek için multipleks polimeraz zincir reaksiyonu analizi yapıldı. İzolatlar arasındaki klonal ilişki Pulsed-Field jel elektroforezi (PFGE) metodu ile araştırıldı.

Bulgular: vanA geni, 86 enfeksiyon ve 64 kolonizasyon izolatı olmak üzere 150 izolatta belirlendi. Bu izolatlar, PFGE analizinde 24 ana kümeye (A-Y) ayrıldı. Her iki grup için en yaygın virülans geni infeksiyon ve kolonizasyon izolatlarında sırasıyla esp geni (% 55.8 ve% 56.25) olarak belirlendi. İkinci sırada ise iki grup arasında istatistiksel olarak farklılık gösteren hyl (% 51.1 ve% 35.9) bulunmakta idi. 

Sonuç: Virülans genlerinin dağılımı ve enfeksiyon ve kolonizasyon gruplarının gen kombinasyonları incelendiğinde, iki grupta küme dağılımının istatistiksel olarak anlamlı olmadığı tespit edilmekle beraber, hyl geninin infeksiyon izolatlarında daha sık görüldüğü tespit edilmiştir.

References

  • Eliopoulos GM, Gold HS. Vancomycin-Resistant Enterococci: Mechanisms and Clinical Observations. Clin Infect Dis. 2001; 33: 210-219.
  • Sujatha S, Praharaj I. Glycopeptide Resistance in Gram-Positive Cocci: A Review. Interdis Persp Infect Dis. 2012; 781679: 10.
  • Mundy LM, Sahm DF, Gilmore. Relationships between Enterococcal Virulence and Antimicrobial Resistance. Clin Microbiol Rev. 2000; 13: 513–522.
  • Çetinkaya Y, Falk P, Mayhall CG. Vancomycin-Resistant Enterococi. Clin Microbiol Rev. 2000; 13: 686-707.
  • Leclercg R, Courvalin P. Resistance to Glycopeptides in Enterococci. Clin. Infect Dis. 1997; 24: 545-56.
  • Van der Heijden KM, Van der Heijden IM, Galvao FH, Lopes CG, Costa SF, Abdala E, D’Albuquerque LA, Levin AS. Intestinal Translocation of Clinical Isolates of Vancomycin Resistant Enterococcus faecalis and ESBL-Producing Escherichia coli in a Rat Model of Bacterial Colonization and Liver Ischemia/Reperfusion Injury. PLOS One. 2014; 9: e108453.
  • Higuita NIA, Huycke MM. Enterococcal Disease, Epidemiology, and Implications for Treatment. In: Gilmore MS, Clewell DB, Ike Y, et al. editors. Enterococci: From Commensals to Leading Causes of Drug Resistant Infection. Boston: Massachusetts Eye and Ear Infirmary. 2014 Available from: http://www.ncbi.nlm.nih.gov/books/NBK190429/.
  • Terkuran M, Erginkaya Z, Ünal E, Güran M, Kızılyıldırım S, Ugur G, Köksal F. The Relationship between Virulence Factors and Vancomycin Resistance among Enterococci collected from food and human samples in Southern Turkey. Ankara Üniv Vet Fak Derg. 2014; 61: 133-140.
  • Togay OS, Keskin AÇ, Açık L, Temiz A. Virulence Genes, Antibiotic Resistance and Plasmid Profiles of Enterococcus faecalis and Enterococcus faecium from Naturally Fermented Turkish Foods J Appl. Microbiol. 2010; 109: 1084-1092
  • Garcia-Migura L, Pleydell E, Barnes S, Davies RH, Liebana E. Characterization of Vancomycin-Resistant Enterococcus faecium Isolates from Broiler Poultry and Pig Farms in England and Wales. J Clin Microbiol 2005; 43: 3283–3289.
  • Karakeçe E, Çiftçi IH., Aşık G. Vankomisin Dirençli Enterokoklarda Direncin Moleküler Yöntemlerle Araştırılması. Ankem Derg. 2013; 27: 135-139.
  • Corso AC, Gagetti PS, Rodrı´Guez MM, Melano RG, Ceriana PR, Faccone DF, Galas M.F. Molecular Epidemiology of Vancomycin-Resistant Enterococcus faecium in Argentina. Inter J Inf Dis. 2007; 11: 69-75.
  • Mirzaei B, Farivar TN, Juhari P, Mehr MA, Babaei R. Investigation of the Prevalence of vanA and vanB Genes in Vancomycin Resistant Enterococcus (VRE) by TaqMan RealTime PCR Assay. J Mic Inf Dis. 2013; 3: 192-198.
  • Domingo MC, Huletsky A, Giroux R, Boissinot K, Picard FJ, Lebel P, Ferraro MJ, Bergeron MG. High Prevalence of Glycopeptide Resistance Genes vanB, vanD, and vanG not Associated With Enterococci in Human Fecal Flora. Antimicrob Agents Ch. 2005; 49: 4784–4786.
  • Rice LB, Lakticová V, Carias LL, Rudin S, Hutton R, Marshall SH. Transferable Capacity for Gastrointestinal Colonization in Enterococcus faecium in a Mouse Model. J Infect Dis. 2009; 199(3): 342-349.
  • Leendertse M, Willems RJL, Oei GA, Florquin S, Bonten MJM., Van der Poll M. Intestinal Enterococcus faecium colonization Improves Host Defense During Polymicrobial Peritonitis. J Infect Dis. 2009; 200: 735-744.
  • Diani M, Esiyok OG, Ariafar MN, Yuksel FN, Altuntas EG, Akcelik F. The Interactions between esp, fsr, gelE genes and Biofilm Formation and PFGE analysis of Clinical Enterococcus faecium strains. African J Microbiol Res. 2014; 8: 129-137.
  • Camargo ILBC, Gilmore MS, Darini ALC. Multilocus sequence typing and analysis of putative virulence factors in vancomycin-resistant and vancomycin-sensitive Enterococcus faecium strains isolated in Brazil. Clin Microbiol Infect. 2006; 11: 1130.
  • Çopur ŞS, Şahin F, Göçmen JS. Determination of Virulence and Multidrug Resistance Genes with Polymerase Chain Reaction Method in Vancomycin Sensitive and Resistant Enterococci Isolated from Clinical Samples. Turk J Med Sci. 2016; 46(3): 877-91.
  • Freitas AR, Tedim AP, Novais C, Ruiz-Garbajosa P, Werner G, Laverde-Gomez JA, Cantón R, Peixe L, Baquero F, Coque TM. Global Spread of the hyl(Efm) Colonization-Virulence Gene in Megaplasmids of the Enterococcus faecium CC17 polyclonal subcluster. Antimicrob Agents Chemother. 2010; 54(6): 2660-2665.
  • Vankerckhoven V, Van Autgaerden T, Vael C, Lammens C, Chapelle S, Rossi R, Jabes D, Goossens H. Development of a Multiplex PCR for the Detection of asa1, gelE, cylA, esp, and hyl genes in Enterococci and Survey for Virulence Determinants among European Hospital Isolates of Enterococcus faecium. J Clin Microbiol. 2004; 42: 4473-4479.
There are 21 citations in total.

Details

Primary Language English
Subjects Medical Microbiology
Journal Section Research
Authors

Tülin Güven Gökmen 0000-0001-9673-097X

Togrul Nagiyev 0000-0002-5719-370X

Beril Akçimen This is me 0000-0002-9649-5241

Begüm Kayar This is me 0000-0002-9657-5970

Melda Meral This is me 0000-0002-5628-6154

Fatih Köksal This is me 0000-0003-0790-1525

Publication Date December 29, 2019
Acceptance Date May 12, 2019
Published in Issue Year 2019 Volume: 44 Issue: 4

Cite

MLA Güven Gökmen, Tülin et al. “Determination of Clonal Relationships and Virulence Genes of Vancomycin-Resistant Enterococcus Spp. Isolated from Colonized and Infected Patients”. Cukurova Medical Journal, vol. 44, no. 4, 2019, pp. 1442-9, doi:10.17826/cumj.552522.