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 MUTALIB^1,2* 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

Received: 18 October 2020/Accepted: 9 March 2021

ABSTRAK

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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*Corresponding author; email: sahilah@ukm.edu.my