Yara Örneklerinden İzole Edilen Metisilin’e Dirençli Staphylococcus Aureus Suşlarında Virulans Genlerinin ve Klonal İlişkilerinin Araştırılması

Year 2020, Volume: 3 Issue: 3, 137 - 144, 01.11.2020

Mert Sudağıdan , Samet Uçak , Orhan Yavuz , Mediha Nur Zafer Yurt Behiye Büşra Taşbaşı Elif Esma Acar Veli Cengiz Özalp Ali Aydın , Şöhret Aydemir

Abstract

Amaç: Staphylococcus aureus insanlarda deri ve yara enfeksiyonlarına neden olan en önemli patojen bakterilerdendir. Metisiline dirençli S. aureus (MRSA) suşlarının yaralarda varlığı hastaların tedavisini zorlaştırmaktadır. Çalışmamızda yara sürüntülerinden izole edilen ve MRSA olarak tanımlanan suşların antibiyotik duyarlılıkları, virulans gen içerikleri ve izolatlar arasındaki genetiksel ilişki araştırılmıştır.
Gereç ve Yöntem: Yara örneklerinden izole edilen suşlar biyokimyasal ve moleküler yöntemlerle tanımlanmıştır. Suşların 12 antibiyotiğe karşı duyarlılıkları agar disk difüzyon yöntemi uygulanarak, indüklenebilir klindamisin direnci D-testi ile ve virulans gen içerikleri PZR ile belirlenmiştir. Suşlar arasındaki genetiksel ilişkinin belirlenmesinde PFGE analizi uygulanmış, SmaI enzimi ile elde edilen band profillerinden dendrogram oluşturularak filogenetik analiz yapılmıştır.
Bulgular: Çalışmada 18 MRSA suşu tanımlanmış ve tümünün penisilin, tetrasiklin, rifampin ve gentamisin'e karşı dirençli oldukları bulunmuştur. Buna karşın tüm suşların linezolid, trimetoprim-sülfametoksazol, kloramfenikol ve quinupristin/dalfopristin'e karşı ise duyarlı oldukları tespit edilmiştir. Ayrıca 4 MRSA suşu indüklenebilir klindamisin direnci göstermiştir. PZR çalışmalarında tüm suşların ermA ve spc-ermA genlerini taşıdığı bulunmuştur. Diğer yandan tetK, tetL, ermB, dfrK, vgaC, ermT, msrA ve msrB genleri suşlarda tespit edilememiştir. ileS, mrm ve spc genleri suşların %94’ünde (17/18) ve ermC geni ise suşların %17’sinde (3/18) pozitif bulunmuştur. Sadece MRSA 50B suşu PVL geni taşıdığı tespit edilmiştir. PFGE analizinde izole edilen MRSA suşları arasında %100 ila %69.4 arasında genetiksel benzerlik olduğu filogenetik analiz ile ortaya konmuştur.
Sonuç: Yara sürüntülerinden izole edilen MRSA suşlarının farklı kökenlerden geldiği ve farklı antibiyotik direnci ve gen içeriklerine sahip olduğu tespit edilmiş ve yara enfeksiyonlarının tedavisinde suşların virulans özelliklerinin de değerlendirilmesinin son derece önemli olduğu sonucuna varılmıştır.

Keywords

Yara, Virulans, Metisiline direnç, Staphylococcus aureus

Determination Virulence Genes and Clonal Relationships of Methicillin Resistant Staphylococcus Aureus Strains Isolated from Wound Samples

Abstract

Objective: Staphylococcus aureus is one of the most important pathogenic bacteria that cause skin and wound infections in humans. The presence of methicillin-resistant S. aureus (MRSA) strains in wounds makes the treatment of patients difficult. In this study, the antibiotic susceptibilities, virulence gene contents and genetic relationships among MRSA strains isolated from wounds were investigated.
Materials and Methods: The strains isolated from wound samples were identified by biochemical and molecular methods. The susceptibility of the strains against 12 antibiotics was determined using agar disk diffusion method, inducible clindamycin resistance by D-test and virulence gene contents by PCR. PFGE analysis was applied to determine the genetic relationships among the strains, phylogenetic analysis was performed by creating a dendrogram from the band patterns obtained with SmaI enzyme.
Results: The results showed that 18 MRSA strains were identified and all strains were found to be resistant to penicillin, tetracycline, rifampin and gentamicin. On the other hand, all strains were found to be susceptible to linezolid, trimethoprim-sulfamethoxazole, chloramphenicol and quinupristin/dalfopristin. In addition, 4 MRSA strains showed inducible clindamycin resistance. In PCR experiments, it was found that all strains carry ermA and spc-ermA genes. Whereas, tetK, tetL, ermB, dfrK, vgaC, ermT, msrA and msrB genes were not detected. ileS, mrm and spc genes were found positive in 94% (17/18) of the strains and the ermC gene in 17% (3/18) of the strains. Only MRSA 50B strain was found to carry the PVL gene. Phylogenetic analysis revealed that there was 100% to 69.4% genetic similarity between the MRSA strains in the PFGE analysis.
Conclusion: MRSA strains isolated from wound swabs came from different origins and had different antibiotic resistance and gene contents, and it was concluded that it is extremely important to evaluate the virulence properties of the strains in the treatment of wound infections.

Keywords

Wound, Virulence, Methicillin resistance

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