Sains Malaysiana 42(12)(2013): 1715–1720

Pengesahan bagi Mengesan Bakteria Patogen Makanan Menggunakan

Teknologi Mikroaturan DNA

(Validation of Food-borne Pathogen Detection using DNA Microarray Technology)

JELIN SAWEI, NORRAKIAH ABDULLAH SANI*, AMINAH ABDULLAH

& SAHILAH ABD. MUTALIB

Pusat Pengajian Sains Kimia dan Teknologi Makanan, Fakulti Sains dan Teknologi

Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor D.E. Malaysia

Diserahkan: 30 November 2011/Diterima: 22 Julai 2013

ABSTRAK

Kajian ini dijalankan untuk mengesahkan kemampuan teknologi DNA mikroaturan cip gen OliproTM FoodPATH bagi mengesan bakteria patogen makanan. Sebanyak 9 jenis DNA bakteria patogen makanan telah digunakan iaitu Bacillus cereus, Escherichia coli O157:H7, Staphylococcus aureus, Vibrio cholerae, Vibrio parahaemolyticus, Listeria monocytogenes, Salmonella spp., Shigella spp. dan Campylobacter spp. Sebanyak 36 kombinasi templat DNA bakteria patogen makanan telah digunakan. Pengesahan bagi mengesan bakteria patogen makanan dilakukan dengan menggunakan kaedah reaksi berantai polimerase (PCR) dan penghibridan Southern-blotting di atas cip gen untuk mengesahkan kemampuannya. Keputusan daripada analisis hibridasi di atas cip gen telah dibandingkan dengan hasil gel elektroforesis 2.0% (w/v). Lima saringan diperlukan untuk menghabiskan 36 kombinasi templat DNA bakteria patogen makanan. Setiap saringan, satu cip gen telah digunakan sebagai kawalan negatif tidak diinokulasikan dengan sebarang kombinasi DNA bakteria patogen makanan. Daripada hasil kajian, didapati bahawa semua kombinasi templat DNA bakteria patogen makanan telah dapat dikesan. Cip yang digunakan sebagai kawalan negatif tidak menunjukkan kehadiran DNA. Oleh itu, daripada kajian ini cip gen OliproTM FoodPATH didapati memberikan keputusan yang lebih baik berbanding dengan 2.0% (w/v) gel elektroforesis.

Kata kunci: Bakteria patogen makanan; cip gen OliproTM FoodPATH, mikroaturan DNA

ABSTRACT

This study was conducted to validate the capability of DNA microarray technology of OliproTM FoodPATH Gene Chip in detecting food-borne pathogens. Nine types of food-borne pathogen DNAs were used namely Bacillus cereus, Escherichia coli 0157:H7, Staphylococcus aureus, Vibrio cholerae, Vibrio parahaemolyticus, Listeria monocytogenes, Salmonella spp., Shigella spp. and Campylobacter spp. A total of 36 combinations of DNA templates of food-borne pathogens were used. Polymerase chain reaction (PCR) and Southern-Blotting Hybridization methods were used on the gene chip to validate its capability.  The results of hibridization on gene chip were compared with results from gel electrophoresis of 2.0% (w/v). Five screenings were required to complete the 36 combinations of DNA templates of food-borne pathogen. For each screening, one gene chip was used as negative control, whereby no DNA templates of food-borne pathogen were inoculated. The results showed that all combinations of food-borne pathogen DNA templates were detected. The negative control chip did not show the presence of any DNA. This study showed that OliproTM FoodPATH Gene Chip gave better results than the 2.0% (w/v) gel electrophoresis.

Keywords: DNA microarray; food-borne pathogen; OliproTM FoodPATH gene ghip

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*Pengarang untuk surat-menyurat; email: norra@ukm.my