Sıkma peynirlerinden izole edilen enterokokların tanımlanmasında MALDI-TOF-MS ve 16S rRNA sekanslamasının karşılaştırılması ve izolatların antibiyotik dirençlikleri ile antimikrobiyal aktivitelerinin belirlenmesi

Öz

Amaç: Bu çalışmanın amacı, sıkma peynirlerden izole edilen enterokokların MALDI-TOF-MS ve 16S rRNA sekans analizleri kullanılarak tanımlanması ve bu izolatların antibiyotik dirençleri ile bazı gıda patojenlerine karşı antibakteriyel etkilerinin değerlendirilmesidir.Yöntem: Sıkma peynirlerden elde edilen 84 Enterococcus izolatının MALDI-TOF-MS yöntemi ile tanımlanması yapılmış ve 16S rRNA bölgeleri çoğaltılarak sekanslanmıştır. İzolatların 14 farklı antibiyotiğe karşı dirençlikleri disk difüzyon yöntemiyle incelenmiştir. Ayrıca izolatların çeşitli gıda patojenlerine karşı gösterdikleri antibakteriyel etkileri agar spot testi ile belirlenmiştir. İstatistiksel değerlendirme için varyans analizi ANOVA, F testi ve antibakteriyel sonuçlar arasındaki korelasyonun tespiti için SPSS 22.0.0 yazılımı SPSS Inc., Chicago, ABD kullanılmışır. Bulgular: Seksen-dört izolat 16S rRNA sekans analizi sonucunda; 33 %39.3 E. faecalis, 29 %34.5 E. faecium, 14 %16.7 E. durans, 4 %4.8 E. gallinarum, 3 %3.5 E. casseliflavus ve 1 %1.2 E. thailandicus olarak tanımlanmıştır. Seksen bir izolat MALDI-TOF-MS yöntemi ile aynı tanımlama sonucunu vermiştir %96.43 . İzolatların tanımlanmasında kullanılan iki tanımlama yönteminin arasında anlamlı bir fark bulunmamıştır p>0.05 . Ancak MALDI-TOF-MS yönteminin daha az iş gücü gerektirmesi, ekonomik ve hızlı olması gibi bazı avantajlarının olduğu görülmüştür. İzolatların %83.3’ünün çoklu antibiyotik direnci taşıdığı tespit edilmiştir. İzolatların nalidiksik asit, oksasilin ve streptomisine karşı dirençliklerinin yüksek olduğu saptanmıştır. E. faecalis suşlarının kullanılan antibiyotiklere karşı E. faecium’dan daha düşük duyarlılık gösterdiği bulunmuştur p

Anahtar Kelimeler

• Gıda, Enterococcus, MALDI, sekanslama, antibiyotik direnci

Comparision of two identification methods: MALDI-TOF-MS & 16s rRNA sequencing in enterococci isolated from raw milk cheeses Sikma cheeses and evaluating their antibiotic susceptibility and antimicrobial activity

Abstract

Objective: This study aims to identify enterococci isolated from squeezed cheeses by MALDI-TOF-MS and 16S rRNA sequence analysis and to evaluate antibiotic resistance and antibacterial effects of these isolates against some food pathogens.Methods: Identification of 84 Enterococcus isolates obtained from squeezed cheese was carried out using MALDI-TOF-MS and 16S rRNA gene sequencing. The isolates were tested for resistance to 14 different antibiotics by the disc diffusion method. The antimicrobial effects of isolates against various food pathogens were determined by the agar spot test. SPSS 22.0.0 software SPSS Inc., Chicago, USA was used for the assessment of the correlation between variance analysis ANOVA, F test and antibacterial results.Results: As a result of sequence analysis; 33 39.3% were described to E. faecalis, 29 34.5% to E. faecium, 14 16.7% to E. durans, 4 4.8% to E. gallinarum, 3 3.5% to E. casseliflavus and 1 1.2% to E. thailandicus. Eighty-one isolates gave the same identification result as the MALDI-TOF-MS method 96.43% . The difference between two identification methods has not been found to be statistically significant p>0.05 ; however, MALDI-TOF-MS has some advantages over 16S rRNA sequencing, such as being less labor-intensive, more economical and faster. In total, 83.3% of the strains exhibited multidrug-resistant phenotypes. A high incidence of resistance was detected for nalidixic acid, oxacillin, and streptomycin. E. faecalis isolates were found to show lower sensitivity to antibiotics tested than E. faecium p

Keywords

• Food, Enterococcus, MALDI, sequencing, antibiotic resistance

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