Leyla BENER, Mustafa ERSAL, Berkant İ. YILDIZ

Genetiği Değiştirilmiş Organizmaların Tespiti ve Ölçümünde Kullanılan Farklı Gerçek Zamanlı PCR Kimyasallarının Karşılaştırılması

Gerçek zamanlı kantitatif polimeraz zincir reaksiyonu (q-PCR), hem gen ekspresyonu analizinde hem de rutin Deoksiribo Nükleik Asit (DNA) ölçümünde nükleik asit miktarının belirlenmesi için kullanılan ileri moleküler bir yöntemdir. Gıda ve yem ürünlerinde genetiği değiştirilmiş organizma (GDO) kalıntıları için etiketleme eşiğinin Japonya’da %5, Avrupa Birliği’nde ise %0,9 olduğu göz önüne alındığında, doğru bir ölçüm metodu şarttır. GDO bileşenlerinin tespiti, kesin miktar tayini ve besin matrislerinde eser miktardaki kalıntısının tespit edilmesi q-PCR’da mümkündür. Bu amaçla çeşitli q-PCR kimyasalları kullanılmaktadır. Bunlar; interkalasyon boyaları, primer bazlı kimyasallar ve prob bazlı kimyasallar olarak üç gruba ayrılmaktadır. Marketlerde GDO ürünlerinin artan sayısıyla birlikte, her örnek için gerçekleştirilen analiz sayısı ve bu nedenle analiz maliyetleri artmaktadır. Bunun için GDO çalışmalarında, GDO’ların varlığının miktarını belirlemede hızlı ve ekonomik olan uygulanabilir taramalar yapılabilmesi için geliştirilmiş tespit yöntemlerine ihtiyaç duyulmaktadır. Bu çalışmada, q-PCR kimyaları ekonomikliği, verimliliği ve uygulanabilirliği açısından karşılaştırılmıştır.

The Comparison of Various Real Time PCR Chemistries Used in Detection and Quantification of Genetically Modified Organisms

Real-time quantitative polymerase chain reaction (q-PCR) is an advanced molecular method for determining the amount of nucleic acid in both gene expression analysis and routine Deoxyribo Nucleic Acid (DNA) measurement. An accurate measurement method is essential given that the labelling threshold for genetically modified organisms (GMO) residues in food and feed products is 5% in Japan and 0,9% in the European Union. Determination of GMO components, quantification of exact amount and determination of trace amounts in food matrices are possible in q-PCR. Various q-PCR chemicals are used for this purpose. These; intercalation dyes, primary based, chemicals and probe based chemicals. With the increasing number of GMO products in the grocery stores, the number of analyses performed per sample and thus the cost of analysis increase. For this purpose, in GMO studies, improved detection methods are needed to determine the presence of GMOs in order to perform fast and economically feasible scans. In this study, q-PCR chemistries were compared in terms of cost, efficiency and applicability.

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