Random amplified polymorphic DNA-Polymerase chain reaction (RAPD-PCR) fingerprinting of Escherichia coli o157:h7

Hasan, Nor'aishah (2008) Random amplified polymorphic DNA-Polymerase chain reaction (RAPD-PCR) fingerprinting of Escherichia coli o157:h7. Masters thesis, University of Malaya.

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Abstract

Twenty (n=20) among beef isolates of Escherichia coli O157:H7 were examined for the detection of Shiga- toxin 1 and 2 (stx1 and stx2) genes and characterized using Random Amplified Polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR) fingerprinting. All isolates were obtained from the laboratory of Food Science and Biotechnology, University Putra Malaysia, Serdang, Selangor. In the detection of stx1 and stx2 genes, 14 of isolates (14/20) were positive to stx1 and stx2. Whereas, 5 isolates (5/20) were positive to stx1 and 1 isolate (1/20) was negative by either of stx1 or stx2 genes. Using RAPD-PCR analysis, two oligonucleotides were chosen because they yielded clearly and reproducible band. There were OPAR8 (5’-TGGGGCTGTC-3’) and OPAR20 (5’-ACGGCAAGGA-3’). Subsequently, all 20 isolates of E.coli O157:H7 were subtyped using OPAR8 and OPAR20. Primer OPAR8 produced 8 RAPD-PCR fingerprinting namely P1 to P11. Whereas, OPAR20 produced 16 RAPD-PCR fingerprinting of Q1-Q18. Combination of two primers was analyzed using Unweighted Pair Group Method with Arithmetic mean (UPGMA). Dendogram performed from cluster analysis showed that all the 20 isolates of E.coli O157:H7 differentiated into 20 individual isolates which may suggest the high level of local geographical genetic variation.

Item Type: Thesis (Masters)
Subjects: Q Science > Q Science (General)
Depositing User: MS SITI NUR ATIKAH MOHAMAD RUSDI
Date Deposited: 31 Jul 2013 03:16
Last Modified: 31 Jul 2013 03:16
URI: http://repository.um.edu.my/id/eprint/1104

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