Investigation of the effect of Aflatoxin B1 and Aflatoxin G1 on DNA Hybridization by Using Electrochemical DNA Biosensor

Gültekin Gökçe; Songül Gökçe; Şenay Çetinus

doi:10.17776/csj.544897

EN TR

Investigation of the effect of Aflatoxin B1 and Aflatoxin G1 on DNA Hybridization by Using Electrochemical DNA Biosensor

Abstract

Biomolecular detection has become a current issue together with the increase in electrochemical sensor studies. The effect of Aflatoxin B1 and Aflatoxin G1 on DNA hybridization was analyzed with electrochemically by using pencil graphite electrode. For this purposes; the changes in guanine currents were monitored before and after hybridization occurred between DNA complementary base sequences using differential pulse voltammetry (DPV). In the first step of this work the capture probe was immobilized by wet adsorption onto the surface of PGE optimizing concentration and immobilization time and then hybridization event was performed between capture and its target probe. In the second step the hybridization experiments were repeated using inosine modified capture probe instead of capture probe which used in the first step. As differently from the first step the inosine modified probe was interacted with 2 µg/mL and 4 µg/mL of AFB1 for 10 min before allowing hybridization and the same protocol was repeated for AFG1. The decrease in guanine current after hybridization was the reference for the genotoxic effects of aflatoxins. The decrease in guanine oxidations in the cases of 2 µg/mL and 4 µg/mL AFG1 and 2 µg/mL and 4 µg/mL AFB1 concentrations were found to be 18% and 26 % for AFG1; and 50% and 61% for AFB1, respectively. AFB1 showed the maximum genotoxic effect to DNA hybridization.

Keywords

Aflatoxin B1,Aflatoxin G1,Pencil Graphite Electrode,Hybridization,Differential Pulse Voltammetry

Aflatoksin B1 ve Aflatoksin G1'in DNA Hibridizasyonuna Etkisinin Elektrokimyasal DNA Biyosensörü ile İncelenmesi

Öz

Biyomoleküler analizlerde yeni teknolojilerin gelişmesi, elektrokimyasal sensör alanındaki çalışmaların da katılmasıyla güncel bir konu olmuştur. Bu çalışmada Aflatoksin B1'in (AFB1) ve Aflatoksin G1'in (AFG1) DNA hibridizasyonu üzerindeki etkisi, kalem grafit elektrot (PGE) kullanılarak elektrokimyasal yöntemlerle analiz edildi. Bu amaçla; guanin oksidasyon akımlarındaki değişiklikler, diferansiyel puls voltametrisi (DPV) ile birbirlerini eşleniği olan DNA baz sekansları arasında hibridizasyon oluşmadan önce ve sonra izlendi. Çalışmanın ilk aşamasında, yakalama probu ıslak adsorpsiyonla PGE yüzeyine immobilize edilirken prob derişimi ve adsorpsiyon süresi optimize edildi ve sonra, hedef prop ile hibritleşmeye tabi tutulurken hedef proba ait konsantrasyon ve hibridizasyon süresi optimize edildi. İkinci aşamada, hibridizasyon deneyleri, birinci aşamada kullanılan yakalama probu yerine inosin modifiye edilmiş yakalama probu kullanılarak tekrarlandı. Ancak, ilk aşamadan farklı bir şekilde inosin modifiye edilmiş prob, hibridizasyona tabi tutulmadan önce 10 dakika süre ile 2 µg / mL ve 4 µg / mL AFB1 ile etkileştirildi ve aynı protokol, aynı koşullarda AFG1 için de tekrarlandı. Hibridizasyondan sonra guanin oksidasyon akımındaki azalma, aflatoksinlerin DNA sekanslarının hibridizasyonuna etkisinin bir sonucu olup, AFB1 ve AFGl'in DNA üzerine genotoksik etkisinin bir göstergesidir. Guanin oksidasyon sinyalindeki azalmalar 2 µg/mL ve 4 µg/mL AFG1 için % 18 ve % 26; 2 µg/mL ve 4 µg/mL AFB1 için % 50 ve % 61 olarak bulundu. Sonuç olarak AFB1'in DNA üzerindeki genotoksik etkisi AFG1'e göre çok daha fazla olarak tespit edildi.

Anahtar Kelimeler

Aflatoksin B1,Aflatoksin G1,Kalem Grafit Elektrot,Hibridizasyon,Diferansiyel Puls Voltametrisi

References

[1] Kerman, K., Meric, B., Ozkan, D., Kara., P., Erdem, A. and Ozsoz, M., Electrochemical DNA biosensor for the determination of Benzo[a]pyrene–DNA adducts, Analytica Chimica Acta, 450 (2001) 45-52.
[2] Ozsoz M., Erdem A., Kerman K., Ozkan D., Tugrul B., Topcuoglu N., Ekren H. and Taylan M., Electrochemical genosensor based on colloidal gold nanoparticles for the detection of factor V Leiden mutation using disposable pencil graphite electrodes, Anal Chem., 75 (2003) 2181-2187.
[3] Palecek E., Fojta, M., Tomschik M., and Wang J., Electrochemical biosensors for DNA damage, Biosensors and Bioelectronics, 13 (1998) 621-628.
[4] Del Giallo M.L., Ariksoysal D.O., Marrazza G., Mascini M. and Ozsoz M., Disposable Electrochemical Enzyme-Amplified Genosensor for Salmonella Bacteria Detection, Analytical Letters, 38 (2005) 2509–2523.
[5] Lin Y., Lu F. and Wang J., Disposable Carbon Nanotube Modified Screen-Printed Biosensor for Amperometric Detection of Organophosphorus Pesticides and Nerve Agents, Electroanalysis, 16(1-2) (2004) 145-149.
[6] Sing Virendra V., Recent Advances in Electrochemical Sensors for Detecting Weapons of Mass Destruction. A Review, ELECTROANALYSIS, 28 (5) (2016) 920-935.
[7] Chen, J. H., Zhang, J., Huang, L. Y., Lin, X. H. and Chen, G. N., Hybridization biosensor using 2-nitroacridone as electrochemical indicator for detection of short DNA species of Chronic Myelogenous Leukemia, Biosensors & Bioelectronics, 24 (2008) 349-355.
[8] Marrazza, G., Chianella, I. and Mascini, M., Disposable DNA electrochemical sensor for hybridization detection, Biosensors & Bioelectronics, 14 (1999) 43-51.

[9] Brabec V. and Koudelka J., Oxidation of DNA at carbon electrodes the effect of the quality of the DNA sample, J. Electroanal. Chem., 116 (1980) 793-799.
[10] Erdem A, Meriç B., Kerman K., Dalbastı T. and Özsöz M., Detection of interaction between metal complex indicator and DNA by using electrochemical biosensor, Electroanal., 11 (1999) 1372-1377.
[11] Jelen, F., Erdem A. and Palecek, E., Cyclic voltammetry of echinomycin and its interaction with double-stranded and single-stranded DNA adsorbed at the electrode, Bioelectrochemistry,55 (2002) 165-173.
[12] Özkan D., Karadeniz H., Erdem A., Mascini M. and Özsöz M., Electrochemical genosensor for Mitomycin C- DNA interaction based on guanine signal, J. Pharmaceutical and Biomedical Analysis, 35 (2004) 905-912.
[13] Wang A.H.J., Interactions between anti tumor drugs and DNA, Nucleic Acids and Molecular Biology, 1 (1987) 52-58.
[14] Pohland, A.E., Mycotoxins in review, Food Additives Contaminants, 10, (1993) 17-28.
[15] Gündüz, S Kimyacılar için istatistik, Gazi Kitabevi, Ankara (1998) 89.
[16] Hazır, Z. and Çoksöyler, N., Farklı bölgelerde ve farklı yöntemlerle elde edilen kırmızı biberlerde aflatoksin düzeyleri, Gıda Mühendisliği Kongre ve Sergisi (1998) 479-483.
17] Trucksess, M.W. and Scott, P.M., Mycotoxins in botanicals and dried fruits: a review. Food Additives and Contaminants. 25 (2008) 181–192.
[18] Williams, J.H., Phillips, T.D., Jolly, P.E., Stiles, J.K., Jolly, C.M., Aggerwal, D. Human aflatoxicosis in developing countries: a review of toxicology, exposure, potential health consequences, and interventions, The American Journal Clinical Nutrition. 80 (2004) 1106-1122.
[19] Fernández-Cruz, M.L., Mansilla, M.L. and Tadeo, J.L., Mycotoxins in fruits and their processed products: Analysis, occurrence and health implications, Journal of Advanced Research, 1 (2010) 113–122.
[20] Zinedine, A., González-Osnaya, L., Soriano, J.M., Moltó, J.C., Idrissi and L., Mañes, J. Presence of aflatoxin M1 in pasteurized milk from Morocco. International journal of food microbiology, 114 (2007) 25–29.
[21] Wang, J., Kawde, A. N. , Erdem A. and Salazar, M., Magnetic bead-based label-free electrochemical detection of DNA hybridization, Analyst, 126 (2001) 2020-2025.
[22] Kuralay, F., Erdem, A., Abacı, S. Ozyoruk, H. and Yıldız, A., Poly (vinylferrocenium) coated disposable pencil graphite electrode for DNA hybridization, Electrochem. Commun., 11(6) (2009) 1242-1246.
[23] Smrdel K.S., Seret M., Duh D., Knap N. and Avsic-Zupanc T., Anaplasma phagocytophilum in ticks in Slovenia, Parasitesand Vectors, 3 (2010) 210-215.
[24] Wang J., Cai X., Rivas G. and Shiraishi H., Stripping potentiometric transduction of DNA hybridization processes, Anal. Chim. Act., 326 (1996) 141-145.
[25] Wang, J.S. and Groopman, J.D., DNA damage by mycotoxins, Mutation Research, 424 (1999) 167–181.

Details

Primary Language

English

Subjects

-

Journal Section

Research Article

Authors

Gültekin Gökçe ^*
0000-0003-1676-194X
Türkiye

Songül Gökçe
0000-0001-5556-6771
Türkiye

Şenay Çetinus
0000-0001-5705-6839
Türkiye

Publication Date

June 30, 2019

Submission Date

March 26, 2019

Acceptance Date

May 10, 2019

Published in Issue

Year 2019 Volume: 40 Number: 2

DOI

https://doi.org/10.17776/csj.544897

IZ

https://izlik.org/JA32AG92CW

Cite

RIS / Bibtex

APA

Gökçe, G., Gökçe, S., & Çetinus, Ş. (2019). Investigation of the effect of Aflatoxin B1 and Aflatoxin G1 on DNA Hybridization by Using Electrochemical DNA Biosensor. Cumhuriyet Science Journal, 40(2), 332-339. https://doi.org/10.17776/csj.544897