Sahilah, A.M. and Aishah, H.N. and Noraida, I. and Azuhairi, A.A. (2010) Detection of Shiga Toxin 1 and 2 (stx1 and stx2) genes in escherichia coli O157:H7 isolated from retail beef in Malaysia by multiplex polymerase chain reaction (PCR). Sains Malaysiana, 39 (1). pp. 57-63. ISSN 0126-6039,
Full text not available from this repository.Abstract
Twenty (n = 20) beef isolates of Escherichia coli O157:H7 were examined for the detection of Shiga-toxin 1 and 2 (stx1 and stx2) genes by multiplex polymerase chain reaction (PCR) 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. 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: | Article |
|---|---|
| Funders: | UNSPECIFIED |
| Uncontrolled Keywords: | Escherichia coli O157:H7; multiplex PCR; retail beef; Stx1 and stx2 genes |
| Subjects: | Q Science > Q Science (General) |
| Depositing User: | MR Faizal II H |
| Date Deposited: | 23 Dec 2015 02:54 |
| Last Modified: | 19 Mar 2019 08:43 |
| URI: | http://eprints.um.edu.my/id/eprint/15256 |
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