Development of Gold-nanoparticle Enhanced Biosensor Applications for Yersinia enterocolitica Detection

Abstract

Yersinia enterocolitica (Y. enterocolitica) is the third most common foodborne zoonotic pathogen belonging to the genus Yersinia which is the member of the Enterobacteriaceae family. It can cause infection in humans as a result of water contaminated with caviar feces or as a result of communication with the infected animal. With its ability to withstand low temperatures, it can continue to reproduce in food in the refrigerator. Therefore, fast and sensitive detection of the pathogen is important for health. In this study, it was aimed to develop an immunosensor reinforced with a gold nanoparticle for the detection of Y. enterocolitica using an electrochemical system. Commercially available Y. enterocolitica positive control and antibody were used in the study. Limit of detection was detected 102 cfu/ml using standard sandwich assay and 37 cfu/ml using gold nanomaterial-amplified antibody sensor. The specificity of the immunoassay was determined by the cross reaction rate of non-antibody-specific antigens. With this study, normal and gold-nanoparticle amplified sandwich assays were developed and used for the detection of commercial Y. enterocolitica. In addition, the effect of gold nanoparticles on measurement sensitivity was determined in the antibody based enzymatic sensor. In the study, electrochemical sensor is used and the immunosensor used in the detection of Y. enterocolitica can be combined with many electrochemical biosensors.

Keywords

• Pathogen detection,antibody,biosensor,gold nanoparticle,Yersinia enterocolitica

Yersinia enterocolitica’nın Tespiti için Altın-nanoparçacık ile Güçlendirilmiş Biyosensör Uygulamalarının Geliştirilmesi

Abstract

Yersinia enterocolitica (Y. enterocolitica), Enterobacteriaceae ailesinin bir üyesi olan Yersinia cinsi’ne ait, en yaygın görülen gıda kaynaklı zoonotik patojenlerden üçüncüsüdür. Havyan dışkısı ile kontamine olan kirli sular ile veya enfekte hayvan ile iletişimin bir sonucu olarak, insanlarda enfeksiyona sebep olabilir. Düşük sıcaklıklara dayanabilme özelliği sayesinde, buzdolabında bulunan yiyeceklerde de üremeye devam edebilmektedir. Bu sebep ile patojenin hızlı ve hassas tespiti sağlık açısında önemlidir. Bu çalışmada, elektrokimyasal sistem kullanılarak, Y. enterocolitica’nın tespiti için altın nanopartikül ile güçlendirilmiş bir immünosensörün geliştirilmesi hedeflenmiştir. Çalışmada ticari olarak temin edilen Y. enterocolitica pozitif kontrol ve antikor kullanılmıştır. Standard sandvich assay kullanılarak sensör yüzeye uygulanan immünoassay ile en düşük tespit limiti 102 cfu/ml, altın nanoparçacıklar ile güçlendirilen antikor sensörü ile 37 cfu/ml Y. enterocolitica tampon çözelti içerisinde tespit edilebilmiştir. İmmünoassay’in özgüllüğü, antikora özgül olmayan antijenlerin çapraz reaksiyon oranı ile tespit edilmiştir. Bu çalışma da, standard sandvich assay ve altın nanopartiküller ile güçlendirilmiş sandvich assay prensibi geliştirilmiş ve Y. enterocolitica’nın tespiti için kullanmıştır. Bunun ile birlikte, antikor tabanlı bir enzimatik sensör de, altın nanoparçacıkların ölçüm duyarlılığına etkisi belirlenmiştir. Çalışmada ölçüm için elektrokimyasal sensör cihazı kullanılmış olup, Y. enterocolitica’nın tespitinde kullanılan immünosensör birçok elektrokimyasal biyosensör ile birleştirilebilir.

Keywords

• Patojen tespit,antikor,biyosensör,altın nanopartikül,Yersinia enterocolitica

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