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

Yıl 2020, Cilt: 15 Sayı: 1, 158 - 166, 31.05.2020
https://doi.org/10.29233/sdufeffd.712282

Öz

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.

Destekleyen Kurum

TÜBİTAK

Teşekkür

Çalışma da kullanılan sensör çip ve elektrokimyasal sensör cihazı TÜBİTAK/Bilgem Biyoelektronik ve Biyosensör Grubu tarafından geliştirilmiş olup, grupta bulunan tüm araştırmacılara teşekkür ederim.

Kaynakça

  • [1] B. M. Rosner, D. Werber, M. Höhle, K. Stark, “Clinical aspects and self- reported symptoms of sequelae Yersinia enterocolitica infections in a population-based study, Germany 2009–2010”, BMC Infect. Dis., 13, 1471–2334, 2013
  • [2] EFSA and ECDC, “The European Union summary report on trends and sources of zoonoses, zoonotic agents and food-borne outbreaks in 2017”, EFSA J., 16, e05500, 2018.
  • [3] E. J. Bottone, “Yersinia enterocolitica: Overview and epidemiologic correlates”, Microbes Infect., 1, 323–333, 1999.
  • [4] L. Tudor, I. Togoe, A. Pop, E. Mitranescu, “The Yersinia enterocolitica species tolerance to temperature”, Rom. Biotechnol. Lett., 13, 17–22, 2008.
  • [5] M. K. Amin, and F. A. Draughon, Growth-characteristics of Yersinia enterocolitica in pasteurized skim milk, J. Food Protect., 50, 849–852, 1987.
  • [6] S. Savas, Z. Altıntas, “Graphene Quantum Dots as Nanozymes for Electrochemical Sensing of Y. enterocolitica in milk and Human Serum”, Materials., 12, 2189, 2019.
  • [7] R. S. Tebbs, L. Y. Wong, P. Brzoska, O. V. Petrauskene, “Molecular technologies for Salmonella detection, In Salmonella Distribution, Adaptation, Control Measures and Molecular Technologies”, IntechOpen, London, UK, 2012, pp. 481–504.
  • [8] I. Hochel and J. Skvor, “Characterization of rabbit antibodies for immunochemical detection of Yersinia enterocolitica”, Folia. Microbiol., 52, 511–518, 2007.
  • [9] M. Luciani, M. Schirone, O. Portanti, P. Visciano, G. Armillotta, R. Tofalo, G. Suzzi, L. Sonsini, T. Di Febo, “Development of a rapid method for the detection of Yersinia enterocolitica serotype O:8 from food”, Food Microbiol., 73, 85–92, 2018.
  • [10] A. Hay, E. Macdonald, R.Evans, M. Davidson, “Use of VITEK for surveillance of antibiotic resistance in Escherichia coli in the Scottish Highlands: results over 15 years”, J. Infection., 55, e87–e88, 2007.
  • [11] D. Derong, W. Liu, H. Li, Y. Wang, X. Li, D. Zou, Z. Yang, S. Huang, D. Zhou, L. Huang, J. Yuan, “Survey and rapid detection of Klebsiella pneumoniae in clinical samples targeting the rcsA gene in Beijing, China”, Front. Microbiol., 6, 519, 2015.
  • [12] E. Sheikhzadeh, M. Chamsaz, A. P. F. Turner, E. W. H. Jager, V. Beni, “Label-free impedimetric biosensor for Salmonella typhimurium detection based on poly pyrrole-co-3-carboxyl-pyrrole copolymer supported aptame”,. Biosens. Bioelectron., 80, 194–200, 2016.
  • [13] B.K. Oh, W. Lee, B. S. Chun, Y. M. Bae, W. H. Lee, J.W. Choi, “Surface plasmon resonance immunosensor for the detection of Yersinia enterocolitica”, Coll Surf A., 257–258, 369–374, 2005.
  • [14] Y. M. Bae, B. K. Oh, W. Lee, W. H. Lee, J. H. Choi, “Immunosensor for detection of Yersinia enterocolitica Based on imaging elipsometry”, Anal. Chem., 76, 1799–1803, 2004.
  • [15] H. Wei, Y. Zhao, Y. Bi, H. Liu, Z. Guo, Y. Song, J. Zhai, H. Huang, R. Yang, “Direct detection of Yersinia pestis from the infected animal specimens by a fiber optic biosensor”, Sens. Actuators B. Chem., 123, 204–210, 2006.
  • [16] W. Sun, P. Qin, H. Gao, G. Li, K. Jiao, “Electrochemical DNA biosensor based on chitosan/nano-V2O5/MWCNTs composite film modified carbon ionic liquid electrode and its application to the LAMP product of Yersinia enterocolitica gene sequence”, Biosens. Bioelectron., 25, 1264–1270, 2010.
  • [17] F. Lu, T.L. Doane, J.-J. Zhu, C. Burda, “Gold nanoparticles fordiagnostic sensing and therapy”, Inorganica Chimica Acta., 393, 142-153, 2012.
  • [18] Z. Altintas, Y. Uludag, Y. Gurbuz, I. Tothill, “Development of surface chemistry for surface plasmon resonance based sensors for the detection of proteins and DNA molecules”, Anal. Chim. Acta., 712, 138–144, 2012.
  • [19] S. Savas, Ersoy. A, Y. Gulmez, S. Kılıc, B. Levent, Z. Altıntas, “Nanoparticle Enhanced Antibody and DNA Biosensors for Sensitive Detection of Salmonella”, Materials., 11, 1541, 2018.
  • [20] S. Savas, “Rapid and Sensitive Detection of E. coli by Gold Nanoparticle-Labeled Biosensor”, J. Biotechnol. Strategic. Health Res., 2, 101-107, 2018.

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

Yıl 2020, Cilt: 15 Sayı: 1, 158 - 166, 31.05.2020
https://doi.org/10.29233/sdufeffd.712282

Öz

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.

Kaynakça

  • [1] B. M. Rosner, D. Werber, M. Höhle, K. Stark, “Clinical aspects and self- reported symptoms of sequelae Yersinia enterocolitica infections in a population-based study, Germany 2009–2010”, BMC Infect. Dis., 13, 1471–2334, 2013
  • [2] EFSA and ECDC, “The European Union summary report on trends and sources of zoonoses, zoonotic agents and food-borne outbreaks in 2017”, EFSA J., 16, e05500, 2018.
  • [3] E. J. Bottone, “Yersinia enterocolitica: Overview and epidemiologic correlates”, Microbes Infect., 1, 323–333, 1999.
  • [4] L. Tudor, I. Togoe, A. Pop, E. Mitranescu, “The Yersinia enterocolitica species tolerance to temperature”, Rom. Biotechnol. Lett., 13, 17–22, 2008.
  • [5] M. K. Amin, and F. A. Draughon, Growth-characteristics of Yersinia enterocolitica in pasteurized skim milk, J. Food Protect., 50, 849–852, 1987.
  • [6] S. Savas, Z. Altıntas, “Graphene Quantum Dots as Nanozymes for Electrochemical Sensing of Y. enterocolitica in milk and Human Serum”, Materials., 12, 2189, 2019.
  • [7] R. S. Tebbs, L. Y. Wong, P. Brzoska, O. V. Petrauskene, “Molecular technologies for Salmonella detection, In Salmonella Distribution, Adaptation, Control Measures and Molecular Technologies”, IntechOpen, London, UK, 2012, pp. 481–504.
  • [8] I. Hochel and J. Skvor, “Characterization of rabbit antibodies for immunochemical detection of Yersinia enterocolitica”, Folia. Microbiol., 52, 511–518, 2007.
  • [9] M. Luciani, M. Schirone, O. Portanti, P. Visciano, G. Armillotta, R. Tofalo, G. Suzzi, L. Sonsini, T. Di Febo, “Development of a rapid method for the detection of Yersinia enterocolitica serotype O:8 from food”, Food Microbiol., 73, 85–92, 2018.
  • [10] A. Hay, E. Macdonald, R.Evans, M. Davidson, “Use of VITEK for surveillance of antibiotic resistance in Escherichia coli in the Scottish Highlands: results over 15 years”, J. Infection., 55, e87–e88, 2007.
  • [11] D. Derong, W. Liu, H. Li, Y. Wang, X. Li, D. Zou, Z. Yang, S. Huang, D. Zhou, L. Huang, J. Yuan, “Survey and rapid detection of Klebsiella pneumoniae in clinical samples targeting the rcsA gene in Beijing, China”, Front. Microbiol., 6, 519, 2015.
  • [12] E. Sheikhzadeh, M. Chamsaz, A. P. F. Turner, E. W. H. Jager, V. Beni, “Label-free impedimetric biosensor for Salmonella typhimurium detection based on poly pyrrole-co-3-carboxyl-pyrrole copolymer supported aptame”,. Biosens. Bioelectron., 80, 194–200, 2016.
  • [13] B.K. Oh, W. Lee, B. S. Chun, Y. M. Bae, W. H. Lee, J.W. Choi, “Surface plasmon resonance immunosensor for the detection of Yersinia enterocolitica”, Coll Surf A., 257–258, 369–374, 2005.
  • [14] Y. M. Bae, B. K. Oh, W. Lee, W. H. Lee, J. H. Choi, “Immunosensor for detection of Yersinia enterocolitica Based on imaging elipsometry”, Anal. Chem., 76, 1799–1803, 2004.
  • [15] H. Wei, Y. Zhao, Y. Bi, H. Liu, Z. Guo, Y. Song, J. Zhai, H. Huang, R. Yang, “Direct detection of Yersinia pestis from the infected animal specimens by a fiber optic biosensor”, Sens. Actuators B. Chem., 123, 204–210, 2006.
  • [16] W. Sun, P. Qin, H. Gao, G. Li, K. Jiao, “Electrochemical DNA biosensor based on chitosan/nano-V2O5/MWCNTs composite film modified carbon ionic liquid electrode and its application to the LAMP product of Yersinia enterocolitica gene sequence”, Biosens. Bioelectron., 25, 1264–1270, 2010.
  • [17] F. Lu, T.L. Doane, J.-J. Zhu, C. Burda, “Gold nanoparticles fordiagnostic sensing and therapy”, Inorganica Chimica Acta., 393, 142-153, 2012.
  • [18] Z. Altintas, Y. Uludag, Y. Gurbuz, I. Tothill, “Development of surface chemistry for surface plasmon resonance based sensors for the detection of proteins and DNA molecules”, Anal. Chim. Acta., 712, 138–144, 2012.
  • [19] S. Savas, Ersoy. A, Y. Gulmez, S. Kılıc, B. Levent, Z. Altıntas, “Nanoparticle Enhanced Antibody and DNA Biosensors for Sensitive Detection of Salmonella”, Materials., 11, 1541, 2018.
  • [20] S. Savas, “Rapid and Sensitive Detection of E. coli by Gold Nanoparticle-Labeled Biosensor”, J. Biotechnol. Strategic. Health Res., 2, 101-107, 2018.
Toplam 20 adet kaynakça vardır.

Ayrıntılar

Birincil Dil Türkçe
Konular Yapısal Biyoloji
Bölüm Makaleler
Yazarlar

Sümeyra SAVAŞ 0000-0001-5057-9178

Proje Numarası Biyosensör ve Biyoelektronik Projesi
Yayımlanma Tarihi 31 Mayıs 2020
Yayımlandığı Sayı Yıl 2020 Cilt: 15 Sayı: 1

Kaynak Göster

IEEE S. SAVAŞ, “Yersinia enterocolitica’nın Tespiti için Altın-nanoparçacık ile Güçlendirilmiş Biyosensör Uygulamalarının Geliştirilmesi”, Süleyman Demirel University Faculty of Arts and Science Journal of Science, c. 15, sy. 1, ss. 158–166, 2020, doi: 10.29233/sdufeffd.712282.