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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)


Citation

Abd Mutalib, Sahilah and Hamzah, Nor' Aishah and Ismail, Noraida and Ariffin, Ahmad Azuhairi (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

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.


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Additional Metadata

Item Type: Article
Divisions: Faculty of Medicine and Health Science
Publisher: Penerbit Universiti Kebangsaan Malaysia
Keywords: Escherichia coli O157:H7; Multiplex PCR; Retail beef; Stx1 and stx2 genes
Depositing User: Raja Norazlinda Raja Azenam
Date Deposited: 29 Apr 2014 12:40
Last Modified: 16 Oct 2018 06:56
URI: http://psasir.upm.edu.my/id/eprint/13938
Statistic Details: View Download Statistic

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