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Ortopetik Cerrahi Yaralardaki Staphylococcus aureus İzolatlarında Slime Üretiminin Genotipik ve Fenotipik Olarak Araştırılması

Yıl 2020, Cilt: 9 Sayı: 1, 17 - 26, 10.06.2020

Öz

Bu çalışmada, Mardin Devlet Hastanesi Ortopedi Servisinde yatan ve implant takılı toplam 180 hastanın yara yerlerinden alınan örneklerin 125’inde Staphylococus aureus suşu izole edilirken, 62’sinde ise Staphylococus epidermitis suşu izole edilmiştir. Bu izolatlarda Multiplex PCR ile slime üretiminden sorumlu icA ve icD genlerinin varlığı, Kongo red agar (CRA), standart test tüp (ST) ve mikropleyt (MP) gibi fenotipik yöntemlerle slime üretimleri, disk difüzyon agar ile suşların antibiyotik dirençlerine bakılarak slime (biyofilm) tabakasının direnç mekanizmasındaki rolünün saptanması amaçlanmıştır. İncelenen stafilokok suşlarının CRA, MP ve ST testleri ile slime üretimi sırasıyla %62.4, %57,7 ve %52.9 oranında pozitif bulunurken icA ve icD gen pozitifliği ise %65,6 (82/125) bulunmuştur. İncelenen tüm stafilokoklar ise teikoplanine ile vankomisin antibiyotiklerine karşı duyarlı oldukları belirlendi. Antibiyotiklere karşı direnç oranlarının slime pozitif suşlarında slime negatif suşlara göre daha yüksek olduğu görülürken, sadece penisilin, ampisilin ve eritromisine dirençlilik oranları istatistiksel olarak anlamlı bulundu. Sonuç olarak, icA ve icD genleri bakımından pozitif olan suşlarda antibiyotiklere dirençlilik oranları anlamlı bulundu.

Kaynakça

  • C.A. Aboltins, V. Antoci, S.Bhattacharyya, M. Cross,
 P. Ducheyne, A.A. Freiberg et al., Hip and Knee Section, Prevention, Prosthesis Factors: Proceedings of International Consensus on Orthopedic Infections, The Journal of Arthroplasty 34 (2019) 309-320.
  • M.E. Rupp G.L. Archer, Coagulase-negative staphylococci: pathogens associated with medical progress, Clin. Infect. Dis. 19 (1994) 231-243.
  • Y.H. An, R.J. Friedmann, Concise review of mechanisms of bacterial adhesion to biomaterial surfaces. J Biomed. Mater. Res. 43 (1998) 338-348.
  • T.J. Foster, D. McDevitt, Molecular basis of adherence of staphylococci to biomaterials, In A.L. Bisno and F.A. Waldvogel (ed.), Infection associated with indwelling medical devices, 2nd ed. American Society for Microbiology, Washington, D.C. 
(1994) 31-43.
  • L. Montanaro, C.R. Arciola, Studying bacterial adhesion to irregular or porous surfaces, In Y. H. An and R. J. Friedmann (ed.), Handbook of bacterial adhesion: principles, methods and applications. Humana Press Inc., Totowa, N.J. 
(2000), pp. 331-343
  • L. Montanaro, C.R. Arciola, L. Baldassarri, E. Borsetti, Presence and expression of collagen adhesin gene (cna) and slime production in Staphylococcus aureus strains from orthopaedic prosthesis infections, Biomaterials 20 (1999) 1945-1949.
  • K.H. Schleifer, Gram Positive Cocci. “Bergey’s Manual of Systematic Bacteriology, Vol 1”. Ed. PHA Sneath, NS Mair, ME Sharpe, JG Holt. Williams and Wilkins, Baltimore, 1986
  • M. Drancourt, A. Stein, J.N. Argenson, A. Zannier, G. Curvale, D. Raoult, Oral rifampin plus ofloxacin for treatment of Staphylococcusinfected orthopaedic implants, Antimicrob. Agents. Chem. 37 (1993) 1214-1218.
  • A. Kumar, R. Prasad, Biofilms, J Med. Educ. Res. 8 (2006) 14-17.
  • C.R. Arciola, L. Baldassarri, L. Montanaro, Presence of ica A and ica D genes and slime production in a collection of staphylococcal strains from catheterassociated infections, J Clin. Microbiol. 39 (2001) 2151-2156.
  • R. Yazdani, M. Oshaghi, A. Havayi, R. Salehi, M. Sadeghizadeh, H. Foroohesh, Detection of icaAD gene and biofilm formation in Staphylococcus aureus isolates from wound infections, Iranian J Publ. Health 35 (2006) 25-28.
  • H.W. Buchholz, H. Engelbrecht, Depot effects of various antibiotics mixed with Palacos resins, Chirurg. 41 (1970) 511-515.
  • L.B. Engesaeter, S.A. Lie, B. Espehaug, O. Furnes, S.E. Vollset, L.I. Havelin, Antibiotic prophylaxis in total hip arthroplasty: effects of antibiotic prophylaxis sys- temically and in bone cement on the revision rate of 22,170 primary hip replacements followed 0-14 years in the Norwegian Arthroplasty Register, Acta. Orthop. Scand. 74 (2003) 644-651.
  • C.J. Gutowski, B.M. Zmistowski, C.T. Clyde, J. Parvizi, The economics of using prophylactic antibiotic-loaded bone cement in total knee replacement, Bone Joint. J. 96 (2014) 65-69.
  • M.J. Dunbar, Antibiotic bone cements: their use in routine primary total joint arthroplasty is justified, Orthopedics 32:9 (2009).
  • CLSI. Clinical and Laboratory Standards Institute/NCCLS. Performance Standards for Antimicrobial Susceptibility Testing; Twenty-Fourth Informational Supplement. Wayne: Clinical and Laboratory Standards Institute, 2014
  • D.J. Freeman, F.R. Falkiner, C.T. Keane, New method for detecting slime production by coagulase negative staphylococci, J Clin. Pathol. 42 (1989) 872–874.
  • G.D. Christensen, W.A. Simpson, J.A. Younger, L.M. Baddour, F.F. Barrett, D.M. Melton et al., Adherence of coagulase negative staphylococci to plastic tissue cultures: A quantitative model for the adherence of staphylococci to medical devices, J Clin. Microbiol. 22(6) (1985) 996-06.
  • W.M. Johnson, S.D. Tyler, E.P. Ewan, F.E. Ashton, D.R. Pollard, K.R. Rozee KR, Detection of genes for enterotoxins, exfoliative toxins, and toxic shock syndrome toxin 1 in S. aureus by the polymerase chain reaction. J Clin. Microbiol. 29 (1991) 426-430.
  • Cramton SE, Gerke C, Schnell NF, Nichols WW, Gotz F (1999). The intercellular adhesion (ica) locus is present in Staphylococcus aureus and is required for biofilm formation. Infect Immun., 67: 5427-5433.
  • Vasudevan P, Nair MK, Annamalai T, Venkitanarayanan KS (2003). Phenotypic and genotypic characterization of bovine mastitis isolates of Staphylococcus aureus for biofilm formation. Vet. Microbiol., 92: 179-185.
  • P. Francois, P. Vaudaux, T.J. Foster, D.P. Lew, Host-bacteria interactions in foreign body infections, Infect. Control Hosp. Epidemiol. 17 (1996) 514-520.
  • E. Rufullayev, Ortopedi servisinde gram-negatif etkenlerle oluşan infeksiyonlar, İstanbul Üniversitesi İstanbul Tıp Fakültesi İnfeksiyon Hastalıkları ve Klinik Mikrobiyoloji Anabilim Dalı. 2018 Thesis
  • C. Gerke, A. Kraft, R. Sussmuth, O. Schweitzer, F. Gotz, Characterization of the N-acetylglucosaminyltransferase activity involved in the biosynthesis of the Staphylococcus epidermidis polysaccharide intercellular adhesin, J. Biol. Chem. 273 (1998) 18586-18593.
  • K. Anagnostakos, Therapeutic use of antibiotic-loaded bone cement in the 
treatment of hip and knee joint infections, J. Bone Jt. Infect. 2 (2017) 29-37. 

  • J.T. Mader, J. Calhoun, Osteomyelitis. In: Mandell GL, Douglas RG, Dolin R, eds. Mandell, Douglas, and Bennett's Principles and Practice of Infectious Diseases, New York: Churchill Livingstone Inc. (2000) 1182-1196.
  • X. Wang, I. Sadovskaya, D. Leterme, D. Watier, A. Chokr, Z. Zhu, S. Jabbouri, A comparative study of antibodies against proteins extracted from staphylococcal biofilm for the diagnosis of orthopedic prosthesis–related infections in an animal model and in humans. Diagnostic Microbiology and Infectious Disease 75 (2013) 124–129
  • H. Özsüt, Ortopedik infeksiyonların güncel tedavisi [Özet], Klinik Derg. 18 (2005) 177.

Genotypic and Phenotypic Investigation of Slime Production in Staphylococcus aureus Isolates in Orthopedic Surgical Wounds

Yıl 2020, Cilt: 9 Sayı: 1, 17 - 26, 10.06.2020

Öz

In this study, Staphylococus aureus strain was isolated in 125 of the samples of 180 patients’wounds who were hospitalized in Mardin State Hospital Orthopedic Service and implanted, while Staphylococus epidermitis strain was isolated in 62 of them. In these isolates, it was aimed to determine the presence of ica A and ica D genes responsible for slime production by Multiplex PCR, to determine slime production by phenotypic methods such as Congo red agar (CRA), standard test tube (ST) and microplate (MP) and to determine the role of the slime (biofilm) layer in the resistance mechanism by looking at the antibiotic resistance of the strains with disc diffusion agar. While the CRA, MP and ST tests of the staphylococcus strains examined were found to be respectively 62.4%, 57.7% and 52.9% positive, the ica A and ica D gene positivity was 65.6% (82/125). It was determined that all staphylococci examined were sensitive to teicoplanin and vancomycin antibiotics. While antibiotic resistance rates were higher in slime positive strains than slime negative strains, only penicillin, ampicillin and erythromycin resistance rates were found statistically significant. As a result, resistance to antibiotics was found to be significant in strains positive with regard to ica A and ica D genes.

Kaynakça

  • C.A. Aboltins, V. Antoci, S.Bhattacharyya, M. Cross,
 P. Ducheyne, A.A. Freiberg et al., Hip and Knee Section, Prevention, Prosthesis Factors: Proceedings of International Consensus on Orthopedic Infections, The Journal of Arthroplasty 34 (2019) 309-320.
  • M.E. Rupp G.L. Archer, Coagulase-negative staphylococci: pathogens associated with medical progress, Clin. Infect. Dis. 19 (1994) 231-243.
  • Y.H. An, R.J. Friedmann, Concise review of mechanisms of bacterial adhesion to biomaterial surfaces. J Biomed. Mater. Res. 43 (1998) 338-348.
  • T.J. Foster, D. McDevitt, Molecular basis of adherence of staphylococci to biomaterials, In A.L. Bisno and F.A. Waldvogel (ed.), Infection associated with indwelling medical devices, 2nd ed. American Society for Microbiology, Washington, D.C. 
(1994) 31-43.
  • L. Montanaro, C.R. Arciola, Studying bacterial adhesion to irregular or porous surfaces, In Y. H. An and R. J. Friedmann (ed.), Handbook of bacterial adhesion: principles, methods and applications. Humana Press Inc., Totowa, N.J. 
(2000), pp. 331-343
  • L. Montanaro, C.R. Arciola, L. Baldassarri, E. Borsetti, Presence and expression of collagen adhesin gene (cna) and slime production in Staphylococcus aureus strains from orthopaedic prosthesis infections, Biomaterials 20 (1999) 1945-1949.
  • K.H. Schleifer, Gram Positive Cocci. “Bergey’s Manual of Systematic Bacteriology, Vol 1”. Ed. PHA Sneath, NS Mair, ME Sharpe, JG Holt. Williams and Wilkins, Baltimore, 1986
  • M. Drancourt, A. Stein, J.N. Argenson, A. Zannier, G. Curvale, D. Raoult, Oral rifampin plus ofloxacin for treatment of Staphylococcusinfected orthopaedic implants, Antimicrob. Agents. Chem. 37 (1993) 1214-1218.
  • A. Kumar, R. Prasad, Biofilms, J Med. Educ. Res. 8 (2006) 14-17.
  • C.R. Arciola, L. Baldassarri, L. Montanaro, Presence of ica A and ica D genes and slime production in a collection of staphylococcal strains from catheterassociated infections, J Clin. Microbiol. 39 (2001) 2151-2156.
  • R. Yazdani, M. Oshaghi, A. Havayi, R. Salehi, M. Sadeghizadeh, H. Foroohesh, Detection of icaAD gene and biofilm formation in Staphylococcus aureus isolates from wound infections, Iranian J Publ. Health 35 (2006) 25-28.
  • H.W. Buchholz, H. Engelbrecht, Depot effects of various antibiotics mixed with Palacos resins, Chirurg. 41 (1970) 511-515.
  • L.B. Engesaeter, S.A. Lie, B. Espehaug, O. Furnes, S.E. Vollset, L.I. Havelin, Antibiotic prophylaxis in total hip arthroplasty: effects of antibiotic prophylaxis sys- temically and in bone cement on the revision rate of 22,170 primary hip replacements followed 0-14 years in the Norwegian Arthroplasty Register, Acta. Orthop. Scand. 74 (2003) 644-651.
  • C.J. Gutowski, B.M. Zmistowski, C.T. Clyde, J. Parvizi, The economics of using prophylactic antibiotic-loaded bone cement in total knee replacement, Bone Joint. J. 96 (2014) 65-69.
  • M.J. Dunbar, Antibiotic bone cements: their use in routine primary total joint arthroplasty is justified, Orthopedics 32:9 (2009).
  • CLSI. Clinical and Laboratory Standards Institute/NCCLS. Performance Standards for Antimicrobial Susceptibility Testing; Twenty-Fourth Informational Supplement. Wayne: Clinical and Laboratory Standards Institute, 2014
  • D.J. Freeman, F.R. Falkiner, C.T. Keane, New method for detecting slime production by coagulase negative staphylococci, J Clin. Pathol. 42 (1989) 872–874.
  • G.D. Christensen, W.A. Simpson, J.A. Younger, L.M. Baddour, F.F. Barrett, D.M. Melton et al., Adherence of coagulase negative staphylococci to plastic tissue cultures: A quantitative model for the adherence of staphylococci to medical devices, J Clin. Microbiol. 22(6) (1985) 996-06.
  • W.M. Johnson, S.D. Tyler, E.P. Ewan, F.E. Ashton, D.R. Pollard, K.R. Rozee KR, Detection of genes for enterotoxins, exfoliative toxins, and toxic shock syndrome toxin 1 in S. aureus by the polymerase chain reaction. J Clin. Microbiol. 29 (1991) 426-430.
  • Cramton SE, Gerke C, Schnell NF, Nichols WW, Gotz F (1999). The intercellular adhesion (ica) locus is present in Staphylococcus aureus and is required for biofilm formation. Infect Immun., 67: 5427-5433.
  • Vasudevan P, Nair MK, Annamalai T, Venkitanarayanan KS (2003). Phenotypic and genotypic characterization of bovine mastitis isolates of Staphylococcus aureus for biofilm formation. Vet. Microbiol., 92: 179-185.
  • P. Francois, P. Vaudaux, T.J. Foster, D.P. Lew, Host-bacteria interactions in foreign body infections, Infect. Control Hosp. Epidemiol. 17 (1996) 514-520.
  • E. Rufullayev, Ortopedi servisinde gram-negatif etkenlerle oluşan infeksiyonlar, İstanbul Üniversitesi İstanbul Tıp Fakültesi İnfeksiyon Hastalıkları ve Klinik Mikrobiyoloji Anabilim Dalı. 2018 Thesis
  • C. Gerke, A. Kraft, R. Sussmuth, O. Schweitzer, F. Gotz, Characterization of the N-acetylglucosaminyltransferase activity involved in the biosynthesis of the Staphylococcus epidermidis polysaccharide intercellular adhesin, J. Biol. Chem. 273 (1998) 18586-18593.
  • K. Anagnostakos, Therapeutic use of antibiotic-loaded bone cement in the 
treatment of hip and knee joint infections, J. Bone Jt. Infect. 2 (2017) 29-37. 

  • J.T. Mader, J. Calhoun, Osteomyelitis. In: Mandell GL, Douglas RG, Dolin R, eds. Mandell, Douglas, and Bennett's Principles and Practice of Infectious Diseases, New York: Churchill Livingstone Inc. (2000) 1182-1196.
  • X. Wang, I. Sadovskaya, D. Leterme, D. Watier, A. Chokr, Z. Zhu, S. Jabbouri, A comparative study of antibodies against proteins extracted from staphylococcal biofilm for the diagnosis of orthopedic prosthesis–related infections in an animal model and in humans. Diagnostic Microbiology and Infectious Disease 75 (2013) 124–129
  • H. Özsüt, Ortopedik infeksiyonların güncel tedavisi [Özet], Klinik Derg. 18 (2005) 177.
Toplam 28 adet kaynakça vardır.

Ayrıntılar

Birincil Dil Türkçe
Konular Yapısal Biyoloji
Bölüm Research Article
Yazarlar

Cemil Demir 0000-0002-6365-0196

Songül Çetik Yıldız 0000-0002-7855-5343

Yayımlanma Tarihi 10 Haziran 2020
Gönderilme Tarihi 2 Nisan 2020
Yayımlandığı Sayı Yıl 2020 Cilt: 9 Sayı: 1

Kaynak Göster

IEEE C. Demir ve S. Çetik Yıldız, “Ortopetik Cerrahi Yaralardaki Staphylococcus aureus İzolatlarında Slime Üretiminin Genotipik ve Fenotipik Olarak Araştırılması”, DÜFED, c. 9, sy. 1, ss. 17–26, 2020.


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