Detection of Clostridium difficile and toxin genes in samples of modified atmosphere packaged (MAP) minced and cubed beef meat

Abstract

In this study it is intended to find out the prevalence of Clostridium difficile in modified atmosphere packaged (MAP) minced (n:50) and cubed beef meat samples (n :50); determine the toxin from the isolates and detect the antibiotic susceptibility against metronidazole, vancomycin and clindamycin. C. difficile isolates have been isolated from two (4%) of 50 minced beef meat samples; one (2%) of 50 cubed beef meat samples. All three isolates were confirmed by PCR as C. difficile by the detection of tpi gene. Three out of 5 C. difficile isolates showed toxigenic character, two of them were carried type B (tcdB) toxin genes, one of them harbored type A (tcdA) toxin gene. When antibiotic resistance profile was phenotypically analyzed, only C. difficile type A (tcdA) was found resistant against clindamycin. All isolates were found sensitive to vancomycin and metronidazole. The result of this study demonstrated that C. difficile strains detected in samples of modified atmosphere packaged (MAP) beef meat could be a potential problem to public health

Keywords

• Antibiotic resistance
• Clostridium difficile
• MAP minced-cubed beef meat
• mPCR

Modifiye atmosfer paketli (MAP) sığır kıyma ve kuşbaşı et örneklerinde Clostridium difficile ve toksin genlerinin belirlenmesi

Abstract

Bu çalışmada konvansiyonel yöntem ve multipleks PCR (mPCR) tekniği ile modifiye atmosfer paketli (MAP) sığır kıyma (n:50) ve kuşbaşı (n:50) örneklerinde Clostridium difficile prevalansı, izolatlardaki toksin gen tipinin belirlenmesi ve metronidazol, vankomisin ve klindamisin antibiyotiklerine karşı duyarlılıklarının tespiti amaçlanmıştır. 50 sığır kıyma örneğinin ikisinde (% 4), 50 sığır kuşbaşı örneğinin birinde (% 2), C. difficile saptanmıştır. Multipleks PCR ile yapılan analizlerde, konvansiyonel yöntemle belirlenen toplam beş izolat ile tpi geninin tespiti ile C. difficile olarak doğrulanmıştır. C. difficile izolatlarının toksin gen varlığı değerlendirildiğinde beş izolatın üçü toksijenik karakterde olup ikisinde C. difficile tip B (tdcB), birinde ise C. difficile tip A (tdcA) toksin geni tespit edilmiştir. Fenotipik antibiyotik direnç profili analizinde klindamisine karşı yalnızca C. difficile tip A (tdcA) toksin genini içeren izolat dirençli bulunurken, tüm izolatlar vankomisine ve metronidazole karşı duyarlı bulunmuştur. Çalışma sonucunda modifiye atmosfer paketli (MAP) et örneklerinde tespit edilen C. difficile’nin halk sağlığı için potansiyel bir tehlike olabileceği ortaya konmuştur

Keywords

• Antibiyotik dirençlilik
• Clostridium difficile
• MAP sığır kıyma ve kuşbaşı
• mPCR

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