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2. Master Thesis / Tesis Master
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Title:	Aplikasi teknologi DNA microarray OliproTM FoodPATH gene chip dalam pengesanan bakteria patogen bawaan makanan.
Authors:	Jelin Sawei (P58617)
Supervisor:	Dr. Norrakiah Abdullah Sani
Keywords:	DNA microarray Bakteria patogen Kaedah cip gen DNA microarrays.
Issue Date:	27-Dec-2013
Description:	Kajian ini telah dijalankan untuk mengesahkan kemampuan teknologi DNA microarray iaitu cip gen OliproTM FoodPATH (Olipro Biotechnology Sdn. Bhd.) mengesan sembilan bakteria patogen bawaan makanan ; Bacillus cereus, Campylobacter spp., Escherichia coli O157: H7, Listeria monocytogenes, Salmonella spp., Shigella spp., Staphylococcus aureus, Vibrio cholerae dan V. parahaemolyticus. Terdapat 36 kombinasi DNA genomik bakteria patogen bawaan makanan digunakan. Analisis hasil hibridasi di atas cip gen dibandingkan dengan keputusan daripada 2.0 % (w/v) elektroforesis gel bagi mengesahkan kemampuan mengesan patogen berkenaan. Daripada keputusan yang diperolehi, telah ditunjukkan bahawa kaedah cip gen, kesemua kombinasi DNA genomik bakteria patogen bawaan makanan telah dikesan, dan dapat memberikan keputusan yang lebih tepat dan jelas jika dibandingkan dengan imej yang dipaparkan oleh elektroforesis gel. Sementara itu, perbandingan keupayaan kaedah cip gen dan pemiringan dalam mengesan bakteria patogen bawaan makanan telah dilakukan. Ia melibatkan dua peringkat iaitu, menggunakan kaedah inokulasi bakteria patogen bawaan makanan di dalam makanan sedia dimakan yang steril dan tanpa kaedah inokulasi dalam makanan sedia dimakan yang bukan steril. Persampelan makanan sedia dimakan telah dilakukan di kafeteria Fakulti Sains dan Teknologi. Terdapat empat kategori makanan digunakan iaitu ayam(ayam goreng), ikan dan produk ikan(ikan kembung goreng masak dalam sos), sayur-sayuran (sayur campuran goreng) dan bijirin (nasi goreng). Bagi kaedah inokulasi di dalam makanan sedia dimakan, cip gen telah berjaya mengesan bakteria patogen daripada 108 (100%) sampel makanan sedia dimakan manakala kaedah pemiringan hanya mampu mengesan bakteria patogen bawaan makanan sejumlah 90 (83.3%) sampel. Untuk pengesanan bakteria patogen bawaan makanan di dalam makanan sedia dimakan yang bukan steril pula, empat sampel nasi goreng dan lima sampel ayam goreng telah dikesan mengandungi B. cereus dengan menggunakan cip gen manakala hanya dua sampel nasi goreng serta tiga sampel ayam goreng telah dikesan mengandungi bakteria tersebut melalui kaedah pemiringan. Cip gen juga mampu mengesan dua sampel nasi goreng dan lima sampel sayur campuran goreng, yang telah dicemari oleh S. aureus, manakala kaedah pemiringan hanya mengesan satu sahaja di dalam sampel nasi goreng. Di samping itu, kaedah pemiringan telah memberikan keputusan positif palsu sebanyak 82.9% (6/35). Secara keseluruhannya, kaedah cip gen adalah lebih cepat, spesifik dan tepat dalam pengesanan bakteria patogen bawaan makanan dalam makanan sedia dimakan dibandingkan dengan kaedah pemiringan.,This study was conducted to validate the capability of DNA microarray technology of OliproTM FoodPATH Gene Chip in detecting nine food-borne pathogens namely Bacillus cereus, Campylobacter spp., Escherichia coli O157: H7, Listeria monocytogenes, Salmonella spp., Shigella spp., Staphylococcus aureus, Vibrio cholerae and V. parahaemolyticus. Thirty six combinations of food-borne pathogens DNA genomic were used. Analysis of the result of hibridization on gene chip were compared with the results from gel electrophoresis of 2.0% (w/v) to validate the capability in detecting those pathogens. The results showed that all combinations of food-borne pathogen DNA genomic were detected, and gave more accurate and clear result compared with the image by gel electrophoresis. The ability of plating and gene chip methods in detecting foodborne pathogens were determined. It consisted of two stages, namely, using spiking method of foodborne pathogen in sterile ready-to-eat (RTE) food and without spiking method in non-sterile RTE food. Ready-to-eat food sampling were done at the Faculty of Science and Technology cafetaria. Four categories of RTE foods were used i.e. poultry (fried chicken), fishery products (fried mackerel cooked in a sauce), vegetables (mixed fried vegetables) and cereals (fried rice). For spiking method on sterile RTE foods, gene chip was able to detect pathogens from 108 (100%) RTE food samples while the plating method was only able to detect in 90 (83.3%) RTE food samples. For the detection of foodborne pathogens in non-sterile RTE foods, four from fried rice and five from fried chicken were detected to contain B. cereus using the gene chip while only two from fried rice and three from fried chicken were detected to contain the said microorganism using the plating method. Gene chip was able to detect two from fried rice and five from mixed fried vegetables contaminated with S. aureus while the plating method was only able to detect one in fried rice. In addition, plating methods gave 82.9 % (6/35) false-positive results. These results indicate that the gene chip method is rapid, specific and accurate in detecting foodborne pathogens in RTE foods compared with the plating method.,Master
Pages:	181
Call Number:	QP624.5.D726 J448 2013
Publisher:	UKM, Bangi
Appears in Collections:	Faculty of Science and Technology / Fakulti Sains dan Teknologi

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