Ulaş ACARÖZ, Damla ARSLAN-ACARÖZ, Zeki GÜRLER

Gıdalarda Antibiyotik Kalıntılarının Saptanması için Enzim İmmunoassay Geliştirilmesi

Veteriner hekimlikte tedavi ve koruyucu amaçla çok sayıda farmakolojik madde kullanılmaktadır. En yaygın ve bilinçsiz kullanılanlarının başında ise antibiyotikler yer almaktadır. Antibiyotikler, hastalıkların tedavisinde önemli rol oynamalarına rağmen yanlış kullanımlarına bağlı olarak et, süt, yumurta, bal gibi hayvansal kökenli gıdalarda kalıntı sorununa neden olmaktadır. Antibiyotik kalıntısı içeren gıdalar halk sağlığı açısından tehlike oluşturmaktır. Halk sağlığını koruma amacıyla, Avrupa Birliği hayvansal kökenli gıdalarda farmakolojik aktif maddeler için maksimum kalıntı limitleri belirlemiştir. Günümüzde gıdalarda antibiyotik kalıntılarının tespiti için geliştirilmiş mikrobiyal inhibisyon testleri, immunoassayler ve kromatografik metotlar mevcuttur. Enzim-linked immunosorbent assay (ELISA) metodu laboratuvarlarda yaygın olarak kullanılmakta, güvenilir sonuçlar vermekte ve kısa sürede çok sayıda örnekle çalışma imkanı sunmaktadır. Bu metodun geliştirilmesinde dikkat edilmesi gereken en önemli aşama immunojenin sentezidir. Antibiyotikler küçük moleküler ağırlığa sahip olduğundan genellikle immun yanıt oluşturamazlar. İmmun yanıt oluşturabilmek için antibiyotik yeterli büyüklükte bir proteine bağlanarak immunojen elde edilir. Antibiyotiğin proteine bağlanması çoğunlukla karbodiimid, glutaraldehid ve aktif ester gibi metotlar ile gerçekleştirilir. Hangi metodun kullanılacağı ilgili antibiyotiğin kimyasal yapısına bağlıdır. Sentezlenen immunojen ile deney hayvanları immunize edilerek, spesifik antikorlar elde edilir. Daha sonra metot optimize edilerek çeşitli gıda maddeleri için geliştirilir.

Anahtar Kelimeler:

Hayvansal Kökenli Gıdalar, Antibiyotik Kalıntısı, Enzim-Linked İmmunosorbent Assay, Metot Geliştirme.

Enzyme Immunoassay Development for the Detection of Antibiotic Residues in Foods

In veterinary medicine, a large number of pharmacological agents are used for prophylactic purpose and treatment. Antibiotics are in the first place among most commonly and inappropriately used pharmacological agents. Although antibiotics play a crucial role in the treatment of the illnesses, misuse of them causes residue problem in the food of animal origin such as milk, eggs, meat, and honey. The food containing antibiotic residues endanger public health. To protect the public health, maximum residue limits for pharmacologically active substances in foods of animal origin have been established by the European Union. To detect the antibiotic residue in foodstuff, several methods such as microbial inhibition tests, immunoassays and chromatographic methods have been established. An enzyme-linked immunosorbent assay (ELISA) method is widely used in a laboratory gives reliable results and offers to work with a large number of samples in a short time. The most important step in the development of this method is the immunogen synthesis. Antibiotics are generally not able to stimulate the immune response because of their low molecular weight. To stimulate the immune response, the immunogen is synthesized by conjugating antibiotics to a protein of sufficient size. The conjugation of antibiotic to protein is mostly performed using some methods such as carbodiimide, glutaraldehyde, and the active ester. Conjugation method is chosen depending on the chemical structure of used antibiotics. Specific antibodies are generated by immunizing the experimental animals with the obtained immunogen. Then, methods are optimized and developed for various foodstuff.

Keywords:

Foods of animal origin, Antibiotic residue, Enzyme-linked immunosorbent assay, Method development,

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