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)

Sahilah Abd. Mutalib, H. Nor Aishah, I. Noraida, A. Ahmad Azuhairi

Research output: Contribution to journalArticle

8 Citations (Scopus)

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 (14120) were positive to stx/ 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 PI to PII. Whereas, OPAR20 produced 16 RAPD-PCRfingerprinting 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.

Original languageEnglish
Pages (from-to)57-63
Number of pages7
JournalSains Malaysiana
Volume39
Issue number1
Publication statusPublished - Feb 2010

Fingerprint

Shiga-like toxin 1
Shiga-like toxin 2
Escherichia coli O157
random amplified polymorphic DNA technique
Malaysia
DNA-directed DNA polymerase
beef
polymerase chain reaction
genes
food biotechnology
oligonucleotides
food science
cluster analysis
genetic variation

Keywords

  • Escherichia coli O157:H7
  • Multiplex PCR
  • Retail beef: Stx1 and stx2 genes

ASJC Scopus subject areas

  • General

Cite this

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). / Abd. Mutalib, Sahilah; Aishah, H. Nor; Noraida, I.; Azuhairi, A. Ahmad.

In: Sains Malaysiana, Vol. 39, No. 1, 02.2010, p. 57-63.

Research output: Contribution to journalArticle

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