Sains Malaysiana 51(10)(2022): 3285-3294

http://doi.org/10.17576/jsm-2022-5110-14

Biosensor DNA Voltametri Berasaskan Nanozarah Emas Bersalut Elektrod Bercetak Skrin Karbon untuk Pengesanan DNA Organisma Terubah Suai Genetik (GMO)

(Voltammetric DNA Biosensor based on Gold Nanoparticles-Coated Carbon Screen-Printed Electrode for Genetically Modified Organism (GMO) Detection)

NAHDYA KHAIRANI¹, HAN YIH LAU², ZAMRI ISHAK², LEE YOOK HENG^3,* & LING LING TAN³

¹Jabatan Sains Kimia, Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor Darul Ehsan, Malaysia

²Malaysian Agriculture Research Institute (MARDI), 43400 Serdang, Selangor Darul Ehsan, Malaysia

³Pusat Kajian Bencana Asia Tenggara (SEADPRI), Institut Alam Sekitar dan Pembangunan (LESTARI), Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor Darul Ehsan, Malaysia

Received:15 March 2022/Accepted: 23 May 2022

Abstrak

Genosensor voltametri untuk pengesanan DNA organisma terubah suai genetik (GMO) telah dibangunkan berasaskan nanozarah emas (AuNPs) dan elektrod pes karbon bercetak skrin (SPE). AuNPs telah dipegunkan pada SPE melalui kaedah pertumbuhan perantara benih dan dicirikan dengan mikroskop elektron pengimbasan (SEM). 6-Mercapto-1-heksanol (MCH) telah digunakan untuk membentuk lapisan mono prob DNA rantai tunggal (ssDNA) pada SPE terubah suai AuNPs (AuNPs-SPE). Biosensor GMO telah dioptimumkan secara elektrokimia menggunakan oligonukleotida sintetik melalui teknik voltametri denyutan pembezaan (DPV). Biosensor GMO yang dioptimumkan kemudiannya digunakan untuk menganalisis sampel sebenar bagi pengesanan khusus urutan DNA dalam virus mozek kubis bunga, iaitu gen CaMV 35S. Isyarat penghibridan DNA dipantau berdasarkan arus puncak pengoksidaan penunjuk redoks asid monosulfonat antrakuinona (AQMS) semasa interkalasinya ke dalam dupleks DNA. Biosensor GMO memberi rangsangan secara linear kepada DNA terubah suai genetik (GM) antara 0.1 nM dan 300.0 nM dan had pengesanan diperoleh pada 0.06 nM. Sisihan piawai relatif (RSD) kebolehulangan biosensor GMO dianggarkan pada 6.7-7.8%. Biosensor GMO berasaskan AuNPs mempamerkan prestasi pengesanan DNA yang lebih baik daripada segi julat linear dinamik dan had pengesanan berbanding dengan biosensor GMO elektrokimia yang dilaporkan sebelum ini. Biosensor DNA elektrokimia ini menyediakan satu platform pengesanan pakai buang untuk aplikasi dalam ujian bahan GMO.

Kata kunci: Biosensor DNA; elektrod pes karbon bercetak skrin; nanozarah emas; organisma terubah suai genetik; voltametri denyutan pembezaan

Abstract

A voltammetric genosensor for detection of genetically modified organism (GMO) DNA has been developed based on gold nanoparticles (AuNPs) and screen-printed carbon paste electrode (SPE). The AuNPs was immobilized on the SPE via seed mediated growth method and characterized with scanning electron microscopy (SEM). 6-Mercapto-1-hexanol (MCH) has been used to form a well aligned monolayer of single-stranded DNA (ssDNA) probes on the AuNPs-modified SPE (AuNPs-SPE). The GMO biosensor has been electrochemically optimized using synthetic oligonucleotides via differential pulse voltammetry (DPV) technique. The optimized GMO biosensor was then applied to real samples analysis for specific detection of DNA sequence in cauliflower mosaic virus i.e., CaMV 35S gene. DNA hybridization signal was monitored based on oxidation peak current of anthraquinone monosulfonic acid (AQMS) redox indicator during its intercalation into DNA duplex. The GMO biosensor response linearly to genetically modified (GM) DNA between 0.1 nM and 300.0 nM and the limit of detection was obtained at 0.06 nM. The reproducibility relative standard deviation (RSD) of the GMO biosensor was estimated at 6.7-7.8%. The AuNPs-based GMO biosensor exhibited improved DNA sensing performance with respect to dynamic linear range and detection limit compared to previously reported electrochemical GMO biosensor. The proposed electrochemical DNA biosensor for GMO provides a disposable sensing platform for the application in GMO material testing.

Keywords: Differential pulse voltammetry; DNA biosensor; genetically modified organism; gold nanoparticles; screen-printed carbon paste electrode

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^*Corresponding author; email: leeyookheng@yahoo.co.uk