Yazar "Kaynak, Ahmet" seçeneğine göre listele

Listeleniyor 1 - 4 / 4
  • [ X ]
    Öğe
    A Rapid and Cost-Efficient Technique for Simultaneous/Duplex Detection of Listeria Monocytogenes and Escherichia Coli O157:H7 Using Real Time PCR
    (Wiley, 2016) Kaynak, Ahmet; Sakalar, Ergun
    The increasing industrial interest in easy, economical and reliable methods has led to the development and application of DNA-based methods for the detection of microbial pathogens in food. In the present article, we describe the development of a cost-efficient EvaGreen-based real-time Polymerase chain reaction (PCR) technique for simultaneous and practical detection of Listeria monocytogenes (L. monocytogenes) and Escherichia coli O157:H7 (E. coli O157:H7) in the food matrix. EvaGreen-based simplex and duplex real-time PCR showed that specific PCR products were identified by melting curve analysis, and had a reproducible T-m of 77.000.4C for L. monocytogenes and 85.60 +/- 0.2 for E. coli O157:H7.The absolute detection limit of Eva Green-based simplex and duplex real-time PCR was 0.0001 ng/L for DNA of both pathogens. The relative detection limit of the simplex and duplex technique was less than 10 cells/mL for the two pathogens. Practical ApplicationsThis study illustrates a rapid, effective and cheap identification method to simultaneously monitor two foodborne pathogens in food samples. The specificity and sensitivity of this method can be used to clearly identify these two foodborne pathogens in practical food samples based on the specific genes of the species. Hence, this detection method is applicable to surveillance measures for these two foodborne pathogens in the food production chain. Moreover, this technique is used by food control laboratories in a secure way.
  • [ X ]
    Öğe
    Gıda patojenlerinin tespiti için floresan temelli ekonomik DNA test yöntemlerinin geliştirilmesi
    (Çanakkale Onsekiz Mart Üniversitesi, 2015) Kaynak, Ahmet; Şakalar, Ergün
    Gıdalarda mikrobiyal patojen tespiti için real-time PCR tekniklerinin geniş uygulama alanı bulması, ayrıca güvenilir, hızlı, kolay ve ekonomik metotlara artan endüstriyel ilgi, DNA temelli metotların geliştirilmesine yol açmıştır. Bu tezde, Listeria monocytogenes ve Escherichia coli O157:H7'nin kompleks bir gıda matrisinde pratik ve eş zamanlı tespiti için ekonomik bir PCR tekniğinin geliştirilmesi anlatılmıştır. Bakteriyel DNA Multi-Fast DNA İzolasyon Kiti (GIDAGEN, Türkiye) ile izole edilmiştir. GC içeriği birbirinden oldukça farklı olan yayınlanmış primerlerin kombinasyonu ve değerlendirilmesi ile EvaGreen temelli dubleks real-time PCR sistemi geliştirilmiştir. Öncül dubleks PCR, model gıda sistemlerinde L. monocytogenes ve E. coli O157:H7'ye uygulanmıştır. L. monocytogenes ve E. coli O157:H7 tespiti için sırasıyla hylA ve uidA hedef gen bölgeleri çoğaltılmıştır. EvaGreen temelli simpleks ve dubleks real-time PCR sistemi ile spesifik PCR ürünleri erime eğrisi analizleri ile belirlenmiş ve L. monocytogenes için tekrarlanabilir Tm değeri 77.00±0.4°C iken, E. coli O157:H7'nin tekrarlanabilir Tm değerinin 85.60±0.5°C olduğu gösterilmiştir. Primerlerin spesifitesi 10 tane farklı bakteri suşu kullanılarak test edilmiştir ve L. monocytogenes ve E. coli O157:H7 tespiti için primerlerin spesifik olduğu belirlenmiştir. EvaGreen temelli dubleks ve simpleks real-time PCR için mutlak algılama sınırı iki patojen bakteri DNA'sı için de 0.00001 ng/?l olarak bulunmuştur. Simpleks ve dubleks tekniğin göreceli tesbit limiti her iki patojen için 10 hücre/ml'den düşük bulunmuştur.
  • [ X ]
    Öğe
    Practical Molecular Detection Method of Beef and Pork in Meat and Meat Products by Intercalating Dye Based Duplex Real-Time Polimerase Chain Reaction
    (Taylor & Francis Inc, 2016) Sakalar, Ergun; Kaynak, Ahmet
    In this study, a rapid, specific, and low-cost duplex-detection technique of pork and beef was developed by a real-time polimerase chain reaction assay based on fluorescence. Deoxyribonucleic acid was extracted from variable mixtures of pork and beef in sausage and industrial products to develop the duplex assay using the GIDAGEN((R)) Multi-Fast DNA Isolation Kit. Identification of genomes was accomplished in the same tube by their distinctive melting peak, which was 87.5 degrees C for pork and 80.5 degrees C for beef, respectively. The detection limit of the method was 0.01 ng/mu L deoxyribonucleic acidor 0.001% target pork and beef in sausage. The results showed that the intercalating dye based duplex real-time polimerase chain reaction is a potentially sensitive, reliable, and practical assay for the detection of meat species adulterated with beef and pork.
  • Küçük Resim
    Öğe
    Soft hydrogels for dual use: Template for metal nanoparticle synthesis and a reactor in the reduction of nitrophenols
    (Elsevier Science Bv, 2012) Şahiner, Nurettin; Kaynak, Ahmet; Bütün, Sultan
    A novel hydrogel based on poly(sulfopropylmethacrylate) (p(SPM) with different crosslinking degrees was synthesized and characterized. The prepared hydrogels were for the first time, utilized for in situ metal nanoparticle preparation such as Ni, Co, and Cu and employed as a reaction media in catalytic reduction of 4-nitrophenol (4-NP), and 2-nitrophenol (2-NP) to 4-aminophenol and 2-aminophenol, respectively. The experimental parameters that effect reduction rates such as temperature and the amount of catalyst were investigated. The kinetics of the reduction reaction of nitro compounds under different reaction conditions were investigated to determine the activation parameter. Activation energies were found as 33.86 kJ mol(-1) and 24.96 kJ mol(-1) for 4-NP and 2-NP, respectively. It was found that hydrogel-Cu composites can provide 98% activity even at the end of the 7th repetitive usage. (c) 2011 Elsevier B.V. All rights reserved.