Yoğun Bakım Ünitesinden Alınan Alt Solunum Yolu Örneklerinde Üretilen Acinetobacter baumannii İzolatlarında Antibiyotik Direnci ve İntegron Gen Kasetleri

Abstract

Amaç: Antibiyotik direnç genlerinin bakteriler arasındaki yayılımı, enfeksiyon hastalıklarının tedavisinde ciddi sorunlara yol açmaktadır. Son zamanlarda bu direnç genlerin integronlarda da bulunduğu gösterilmiştir. Bu çalışmada alt solunum yollarından alınan kültürlerde üretilen infeksiyon etkeni olan Acinetobacter baumannii suşlarında sınıf 1, sınıf 2 integron varlığı ve antibiyotik direnç genleri araştırıldı. 

Gereç ve Yöntemler: Çalışmaya, Mart 2014- Mart 2015 tarihleri arasında alt solunum yolu örneklerinden izole edilen toplam 86 A. baumannii suşu dahil edildi. İzolatların tanımlanması ve antibiyogramları konvansiyonel metodlar ve Vitek 2 Compact sistemleriyle yapıldı. Suşlarda integron varlığı, sınıf 1 (intI1) ve sınıf 2 (intI2) integraz bölgeleri için özgül primer çiftleri kullanılarak PCR yöntemiyle araştırıldı. İntegron amplifikasyonunun gerçekleştirildiği tüm örnekler hem klonlanarak hem de PCR ürünü olarak DNA dizi analizine tabi tutuldu. Direnç genleri polimeraz zincir reaksiyonu ile tarandı. 

Bulgular: İzolatlar trakeal aspiratlar (%33.7), bronkoalveoler lavaj (%29,1), balgam (%22.1), bronşiyal yıkama sıvısı / bronşiyal fırçalama (%4.7), transbronşiyal biyopsi (%4.7), plevral sıvı (%3.4) ve akciğer aspirasyonları / apse (%2.3) örneklerinden elde edildi. Tüm izolatlarda imipenem, meropenem, seftazidim, piperasillin-tazobaktam, ampisilin-sulbaktama karşı direnç saptanırken, siprofloksasin, levofloksasin, gentamisin, amikasin ve tigesikline karşı direnç sırasıyla %97.7, %93.0, %60.5, %53.5 ve %9.3 olarak tespit edildi. Kolistin, tüm suşlara karşı tek etkin antibiyotikti. Integraz I ve II gen pozitiflik oranı sırasıyla %45.4 ve %9.3 iken, sınıf 1 ve 2 integron pozitiflik oranı tüm izolatlarda sırasıyla %31.4 ve %8.1 olarak bulundu. blaTEM, blaSHV, blaCTX-M1, blaCTX-M2, blaOXA23, blaOXA40, blaOXA58 sırasıyla %70.9, %2.3, %4.7, %7.0, %95.3, %5.8 ve %3.5 bulundu.

Sonuç: A. baumannii infeksiyonlarında çoklu ilaç direnci olan suşların ve integron direnç genlerinin birlikte artışı, antibiyotik tedavi stratejilerinin başarısızlığına yol açacaktır.

Keywords

• Acinetobacter baumannii
• Antibiyotik direnç genleri
• alt solunum yolu örnekleri
• integron

ANTIBIOTIC RESISTANCE AND INTEGRON GENE CASSETTES IN ACINETOBACTER BAUMANNII ISOLATES PRODUCED IN LOWER RESPIRATORY TRACT SAMPLES TAKEN FROM THE INTENSIVE CARE UNIT

Abstract

Objective: The spread of antibiotic resistance genes among bacteria causes serious problems in the treatment of infectious diseases. Recently, it has been shown that these resistance genes are also found in integrons. This study investigated the class 1, 2 integrons and the antibiotic-resistant genes in the infection agent Acinetobacter baumannii produced in respiratory tract samples.

Material and Methods: The study involved 86 A. baumannii strains isolated from lower respiratory tract samples collected between March 2014 and March 2015 in the Intensive Care Units. Identification and antibiograms of the isolates were made using conventional methods and Vitek 2 Compact systems. The presence of integrons in strains was investigated by polymerase chain reaction method using specific primer pairs for class 1and class 2 integrase regions. All samples in which integron amplification was carried out were subjected to DNA sequence analysis both by cloning and as a PCR product. The resistance genes were screened by polymerase chain reaction. 

Results: Isolates were obtained from specimens including tracheal aspirates (33.7%), bronchoalveolar lavage (29.1%), sputum (22.1%), bronchial washing fluid/bronchial brushing (4.7%), transbronchial biopsy (4.7%), pleural fluid (3.4%) and lung aspirates/abscess (2.3%). While resistance to imipenem, meropenem, ampicillin-sulbactam, ceftazidime, and piperacillin-tazobactam was determined in all isolates, resistance rates to ciprofloxacin, levofloxacin, gentamicin, amikacin and tigecycline were 97.7%, 93.0%, 60.5%, 53.5%and 9.3%, respectively. Colistin was the only antibiotic to which all strains were susceptible. The positivity rates of integrase I and II genes were 45.4%and 9.3%, respectively. Class 1 and 2 integron positivity rates in all isolates were 31.4%and 8.1%, respectively. blaTEM, blaSHV, blaCTX-M1, blaCTX-M2, blaOXA23, blaOXA40, blaOXA58 were 70.9%, 2.3%, 4.7%, 7.0%, 95.3%, 5.8%and 3.5%, respectively. 

Conclusion: Increasing association of multi drug resistant strains and integron resistant genes in A. baumannii infections will lead to the failure of antibiotic treatment strategies.

Keywords

• Acinetobacter baumannii
• Lower respiratory tract specimens
• Integron
• Antibiotic-resistant genes

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