Dondurulmuş Deniz Ürünlerinde Vibrio spp.’nin İlmiğe Dayalı İzotermal Amplifikasyon (Loop Mediated Isothermal Amplification, LAMP) Yöntemiyle Belirlenmesi

Yıl 2022, , 14 - 19, 30.06.2022

Yusuf Doğruer , A. Ezgi Telli , Nihat Telli , Yusuf Biçer

https://doi.org/10.47027/duvetfd.1095611

Öz

Bu çalışmada, deniz ürünleri kaynaklı enfeksiyonlarda en yaygın etkenlerden olan Vibrio spp.’nin yüksek risk grubunu oluşturan Vibrio parahaemolyticus, V. vulnificus ve V. cholerae’nın yüksek özgüllük, verimlilik ve hızlılık içeren yeni bir yöntem olan İlmiğe Dayalı İzotermal Amplifikasyon (Loop Mediated Isothermal Amplification, LAMP) yöntemiyle tespit edilmesi amaçlandı.
Araştırma materyali olarak, süpermarketler ve balık hallerinde satışa sunulan dondurulmuş deniz ürünleri (hamsi filetosu, bütün hamsi, dondurulmuş balık köfte, ahtapot, karides, kalamar, akivades midyesi, midye eti, kabuklu midye, uskumru filetosu, sardalya, surimi çubuk, vongole midyesi, çıtır kalamar halkaları, kaya levreği n = 212) toplandı. Klasik kültür yöntemiyle örneklerin izolasyonu için ISO / TS 21872-1: 2007 ve ISO 21872-2: 2007 prosedürleri kullanıldı. Şüpheli izolatlardan DNA ekstraksiyonu gerçekleştirildi. Daha sonra izolatlar Vibrio spp. için gyrB1 gen bölgesinin amplifikasyonu ile genus spesifik PCR reaksiyonu kullanılarak doğrulandı. Pozitif örneklerde turbidite bazlı Real-Time LAMP gerçekleştirildi. Bu amaçla, sırasıyla V. parahaemolyticus, V. vulnificus ve V. cholerae'nin saptanması için toxR, vvHA ve ompW hedef gen bölgeleri için tasarlanmış LAMP primer setleri kullanıldı.
Sonuç olarak, klasik kültür metodu kullanılarak analiz edilmiş 212 (% 16,98) örnekten 36'sının Vibrio spp. ile kontamine olduğu tespit edildi. LAMP reaksiyonu sonuçlarına göre, Vibrio spp. pozitif izolatların içerisinde V. parahaemolyticus ve V. vulnificus'un oranı sırasıyla 13/36 (% 36.1) ve 2/36 (% 5.5) olarak tespit edildi. Analiz edilen örneklerin hiçbirinde V. cholerae tespit edilmedi. Bu bağlamda, dondurulmuş deniz ürünlerinde V. parahaemolyticus ve V. vulnificus gibi patojenik suşların varlığı halk sağlığı açısından riskli olarak kabul edildi.

Anahtar Kelimeler

Dondurulmuş deniz ürünleri, LAMP, Vibrio spp.

Destekleyen Kurum

S.Ü. BAP KOORDİNATÖRLÜĞÜ

Proje Numarası

15401068

Teşekkür

Araştırmayı 15401068 numaralı proje ile destekleyen Selçuk Üniversitesi Bilimsel Araştırma Koordinatörlüğü’ne teşekkür ederiz. Araştırma özeti “3rd International Congress on Advances in Veterinary Science and Technics (ICAVST)” Kongresi’nde sözlü bildiri olarak sunulmuştur.

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