Electrochemical DNA biosensor for detection of pork (Sus scrof) using screen printed carbon-reduced graphene oxide electrode

Abstract

The identification of pork in foodstuff is critical regarding the counterfeiting of meat and kosherness, which is a particular concern for certain religions. In this study, we developed an electrochemical detection method of pork DNA without the use of DNA amplification by using screen printed carbon-reduced graphene oxide (SPC-RGO) electrode. The probe DNA of CytB gene of S. scrofa mtDNA was immobilized on the SPC-RGO surface by passive adsorption. Differential pulse voltammetry (DPV) was used to characterise the probe-target DNA hybridisation based on the target's guanine oxidation signal. The Placket-Burman and Box Behnken designs were used to select the factors that influence the hybridisation of probe-target DNA and to optimise each parameter. The following findings regarding the several factors that influence the hybridisation process and optimum condition were obtained: 5.0 µg/ml of probe DNA, 6.0 min of immobilisation time of probe DNA, 20.0 min of probe-target hybridisation time, a scan rate at 0.5 V/s, the pulse amplitude at 50.0 mV, and the washing time of the electrode being as long as 40 s. The limit of detection was obtained at 1.76 µg/ml for the linear range of 0-10.0 µg/ml target DNA while the relative standard deviation (RSD) was 2.25%. The DNA biosensor was tested on the isolated DNA samples from pork, chicken and beef while the voltammetry response reveals that it can distinguish the samples. These results indicate that the proposed electrochemical DNA biosensor has the potential to develop the detection method of pork content in the food samples.

Cyclic voltammetric determination of capsaicin by using electrochemically deposited tin and reduced graphene oxide on screen-printed carbon electrodes

“…5. This mechanism was proposed in the previous report [18]. The electrochemical oxidation of capsaicinoids is an irreversible step.…”

“…Alternate approach for capsaicin determination could be done via electrochemistry due to its advantages, such as high efficiency, good reproducibility, greater linear response range, and easy operation. Among electrochemical techniques, cyclic voltammetry is the most widely used to study the electrochemical behavior of electroactive species by using biosensor [17,18] or non-biosensor [19].…”

Modifikasi Screen Printed Carbon Electrode Menggunakan Cerium dan Optimasi Kondisi Percobaan Biosensor Elektrokimia untuk Deteksi DNA Mitokondria Sus Scrofa

“…Beberapa penelitian telah digunakan untuk menganalisis urutan spesifik DNA mitokondria babi (Bielikova, et al, 2010;Erwanto, et al, 2011;Maryam, et al, 2016). Sedangkan penelitian yang telah dilakukan untuk mengidentifikasi kandungan babi menggunakan biosensor yaitu nanobioprobe untuk identifikasi daging babi di burger (Ali, et al, 2012a), optik nanobiosensor untuk deteksi dan kuantifikasi DNA babi dalam campuran daging (Ali, et al, 2012b), dan deteksi DNA babi dengan kompleks [Ru(bpy)2(PIP)] 2+ (Halid, et al, 2014), biosensor voltammetri menggunakan SPCE-reduced graphene oxide electrode (Hartati, et al, 2018), biosensor voltammetri menggunakan SPCE-AuNP (Screen Printed Carbon Electrode-Nanopartikel Emas) (Hartati, et al, 2019).…”

“…Penelitian sebelumnya, nilai LoD dari biosensor DNA elektrokimia menggunakan grafena sebagai pemodifikasi SPCE didapatkan sebesar 1,76 ng/µL (Hartati, et al, 2018), DNA swine dalam daging sapi olahan didapatkan masing-masing sebesar 6,0 ng/µL dan 4,0 ng/µL menggunakan nanopartikel emas sebagai sensor kolometri (Ali, et al, 2012a;Ali, et al, 2012b), dan DNA pork dalam bakso didapatkan sebesar 0,58 µg/mL menggunakan AuNP-biokonjugat pada elektrode SPCE-emas (Hartati, et al, 2019).…”

Biosensor DNA Elektrokimia untuk Deteksi Makanan Mengandung Babi (Sus scrofa) Menggunakan Screen Printed Carbon Electrode Termodifikasi Emas

“…Hasil pengukuran menunjukkan batas deteksi 0,58 µg/mL dari rentang konsentrasi DNA komplementer 0 -5 µg/mL. Sementara itu, penggunaan SPCE termodifikasi grafen menghasilkan batas deteksi 1,76 µg/mL untuk range konsentrasi 0 -10 µg/mL (Hartati et al, 2020). Kedua penelitian ini menggunakan DNA probe yang dilaporkan oleh Ali et al (2012).…”