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Sains Malaysiana 54(8)(2025): 1957-1971
http://doi.org/10.17576/jsm-2025-5408-07
Biosensor DNA
Elektrokimia Berasaskan Nanokomposit Silika-Emas untuk Penentuan Ultrasensitif E.
coli Patogen dalam Sayuran dan Air Persekitaran
Electrochemical DNA Biosensor Based
on Silica-Gold Nanocomposite for Ultrasensitive Quantitation of Pathogenic E.
coli in Vegetable and Environmental Water
LING LING TAN1, DEDI FUTRA2,3,*
& LEE YOOK HENG2
1Pusat Kajian Bencana Asia Tenggara (SEADPRI), Institut Alam Sekitar
dan
Pembangunan
(LESTARI), Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor, Malaysia
2Jabatan Sains Kimia, Fakulti Sains dan Teknologi, Universiti
Kebangsaan Malaysia,
43600 UKM
Bangi, Selangor, Malaysia
3Department of Chemistry Education, Faculty of Education, Universitas
Riau, Kampus Binawidya KM 12.5 Pekanbaru, 28131, Riau, Indonesia
Diserahkan: 27 Disember 2024/Diterima: 23 Jun
2025
Abstrak
Escherichia coli lazimnya digunakan sebagai organisma penunjuk untuk menilai kualiti air
dan pencemaran patogen enterik di perairan. Mengesan bakteria najis, seperti E.
coli O157:H7 yang mempunyai ciri virulensi yang
akan menyebabkan kesan patogen pada manusia adalah lebih membimbangkan orang
biasa. Di sini, biosensor DNA elektrokimia ultrasensitif berasaskan nanokomposit silika-emas (Si-Au) telah disediakan
untuk penentuan khusus jujukan DNA E. coli dalam
sampel sayuran segar dan air daripada pelbagai sumber, contohnya sungai,
laut, paip dan air mineral. Elektrod pes karbon
bercetak skrin (SPCE) telah diubahsuai dengan lapisan nanozarah emas
(AuNPs) untuk menggalakkan pemindahan elektron secara langsung antara label
oligonukleotida antrakuinona elektroaktif dan elektrod SPCE. Nanosfera silika terubah
suai dengan kumpulan berfungsi amina (SiNPs) telah disintesis melalui teknik
sonikasi satu langkah yang mudah dan dijerap secara fizikal pada SPCE terubah
suai AuNPs untuk membentuk elektrod nanokomposit Si-Au (Si-Au-SPCE). Biosensor
DNA elektrokimia terbangun mampu mengesan kuantiti DNA sasaran E. coli pada paras rendah dalam julat kepekatan 1.0×10-16-1.0×10-9 M cDNA dengan had pengesanan (LOD) pada 0.13 aM. LOD yang dilaporkan dalam
kajian ini adalah sekurang-kurangnya 10 kali ganda lebih rendah daripada
kebanyakan biosensor DNA elektrokimia yang dilaporkan dalam kajian lepas
setakat ini. Kuantifikasi DNA E. coli hanya memerlukan kira-kira 30
minit dan biosensor DNA menunjukkan kestabilan operasi selama 2 bulan.
Nanokomposit Si-Au boleh berfungsi sebagai bahan termaju yang berpotensi untuk
biosensor DNA berprestasi tinggi seperti yang dibuktikan melalui kajian
kuantitatif kepekatan DNA E. coli dalam sampel sayuran mentah dan air
persekitaran dengan hampir 100% perolehan semula kepekatan cDNA E. coli yang ditambahkan dalam sampel.
Kata kunci: Air
persekitaran; biosensor DNA elektrokimia; Escherichia coli; nanosfera
silika; nanozarah emas
Abstract
Escherichia coli is commonly used as an indicator organism for
assessing water quality and enteric pathogenic contamination in waters.
Detecting fecal bacteria, such as E. coli O157:H7 that had
acquired virulence characteristics, which would cause pathogenicity in humans is of more concern to
the average people. Herein, an
ultrasensitive electrochemical DNA biosensor based on silica-gold (Si-Au)
nanocomposite was prepared for sequence-specific detection of E. coli DNA in fresh vegetable and water samples from various sources (river, marine,
tap, and mineral waters). The screen-printed carbon paste electrode (SPCE) was
modified with a layer of gold nanoparticles (AuNPs) to promote direct electron transfer between
the electroactive anthraquinone oligonucleotide label and the SPCE electrode. Animated-silica
nanospheres (SiNPs) were synthesized via a facile single-step sonication
technique, and physically absorbed onto the AuNPs-modified SPCE to form Si-Au
nanocomposite electrode (Si-Au-SPCE). The electrochemical DNA biosensor was
capable of low levels of E. coli target DNA quantitation in the concentration range of 1.0×10-16-1.0×10-9 M cDNA, with a limit of detection (LOD) at 0.13 aM. The LOD reported in this
work is at least 10 times better than many reported electrochemical DNA
biosensors in the literature so far. The quantification of E. coli DNA requires only about 30 min and the DNA biosensor showed
operational stability for 2 months. The Si-Au nanocomposite could serve as a
promising material for high-performance DNA biosensor as proven through the
quantitation of E. coli DNA
concentration in raw vegetable and water samples with close to 100% recoveries
of the spiked E. coli cDNA
concentration.
Keywords: Electrochemical DNA biosensor; environmental
waters; Escherichia coli; gold
nanoparticles; silica nanospheres
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*Pengarang untuk
surat-menyurat; email: dedifutra@lecturer.unri.ac.id
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