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Sains Malaysiana 50(11)(2021): 3275-3284
http://doi.org/10.17576/jsm-2021-5011-11
Pengasingan
dan Pengesanan Asid Deoksiribonukleik (DNA) Spesies Haiwan menggunakan
Multipleks Tindak Balas Rantaian Polimerase (PCR) daripada Minyak Sapi
(Isolation
and Identification of Deoxyribonucleic (DNA) of Animal Species by Multiplex
Polymerase Chain Reaction (PCR) from Ghee)
NURUL HANISAH NOOR AZLI1, SAHILAH ABD MUTALIB1,2*
HASLANIZA HASHIM1, MAARUF ABD GHANI1 & MOHD IZHAR
ARIFF MOHD. KASHIM3
1Department of Food Sciences, Faculty of Science and
Technology, Universiti Kebangsaan Malaysia
43600 UKM Bangi, Selangor Darul
Ehsan, Malaysia
2Innovation Centre for Confectionery Technology (MANIS),
Faculty of Science and Technology
Universiti Kebangsaan Malaysia,
43600 UKM Bangi, Selangor Darul Ehsan, Malaysia
3Centre of Shariah, Faculty of Islamic Studies, Universiti
Kebangsaan Malaysia, 43600 UKM Bangi, Selangor Darul Ehsan, Malaysia
Diserahkan: 18 Oktober
2020/Diterima: 9 Mac 2021
Kajian ini dijalankan untuk mengesan asid deoksiribonukleik
(DNA) spesies haiwan dalam campuran minyak sapi menggunakan teknik tindak balas
rantaian polimerase (PCR) multipleks. Kebolehdapatan DNA daripada produk
berminyak membolehkan analisis PCR dijalankan dan sekaligus boleh menentukan
kehadiran spesies haiwan produk tersebut dengan menggunakan primer haiwan yang
khusus-spesies. Gen sasaran adalah DNA mitokondria (mtDNA) Sitokrom oksidase II
(COII) untuk menentukan lembu, manakala mtDNA tRNA-ATP8 dan DNA nuklear (nDNA)
Elemen Nuklear Berselang pendek (SINE) untuk menentukan babi. Terdapat satu
kawalan (S1) dan tiga sampel campuran minyak sapi dengan gelatin babi ditambah
pada kepekatan berbeza iaitu 1%, 3% dan 5% (w/v) (S2, S3 dan S4). Analisis PCR
simpleks menggunakan primer spesies-spesifik yang disebutkan, berjaya
mengamplifikasi DNA lembu dengan amplikon bersaiz 165 bp (COII) dan DNA babi
dengan amplikon 212 bp (tRNA-ATP8) dan 161 bp (SINE). Manakala, teknik PCR
multipleks yang telah dioptimumkan menggunakan primer mtDNA (tRNA-ATP8) dan
nDNA (SINE) untuk menentukan kehadiran haiwan babi, juga menghasilkan amplikon
bersaiz yang sama. Oleh itu, kajian menunjukkan bahawa kaedah PCR multipleks
adalah kaedah yang mudah, ringkas dan pantas untuk menentukan kehadiran DNA
babi di dalam sampel campuran produk minyak sapi selain daripada kaedah PCR
simpleks.
Kata
kunci: Minyak sapi; PCR multipleks; pengasingan; pengenalpastian DNA; primer
khusus-spesies
ABSTRACT
This study was conducted to detect deoxyribonucleic acid
(DNA) of animal species in ghee mixture using polymerase chain reaction (PCR)
technique. The availability of DNA from oily products allows PCR analysis to be
carried out and at the same time can determine the presence of animal species
of the product using species-specific animal primers. The target genes are
mitochondrial DNA (mtDNA) cytochrome oxidase II (COII) to determine bovine,
while tRNA-ATP8 mtDNA and nuclear DNA (nDNA) Short Intermediate Nuclear
Elements (SINE) to determine porcine. There was one control (S1) and three
samples of ghee mixture with porcine gelatine added at different concentrations
of 1%, 3%, and 5% (w/v) (S2, S3, and S4). Simplex PCR analysis using the
species-specific primers mentioned above, successfully implicated bovine DNA
with 165 bp amplicons (COII) and porcine DNA with 212 bp (tRNA-ATP8) and 161 bp
(SINE) amplicons. Whereas, the multiplex PCR technique that has been optimized
using mtDNA (tRNA-ATP8) and nDNA (SINE) primers to determine the presence of
porcine, also produced amplicons of the same size as above. Therefore, studies
show that the multiplex PCR method is an easy, simple and fast method to
determine the presence of porcine DNA in ghee products mixed sample other than
simplex PCR method.
Keywords:
DNA identification; ghee; multiplex PCR; separation; species-specific primer
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*Pengarang untuk surat-menyurat;
email: sahilah@ukm.edu.my
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