Detection of shiga toxin 1 and 2 (stx1 and stx2) genes in escherichia coli 0157: H7 isolated from retail beef in Malaysia by multiplex polymerase chain reaction (PCR)

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 isolate...

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Main Authors:	A.M. Sahilah, H. Nor' Aishah, A. Ahmad Azuhairi
Format:	Article
Language:	English
Published:	Universiti Kebangsaan Malaysia 2010
Online Access:	http://journalarticle.ukm.my/7304/ http://journalarticle.ukm.my/7304/ http://journalarticle.ukm.my/7304/1/03.pdf

id	ukm-7304
recordtype	eprints
spelling	ukm-73042016-12-14T06:43:41Z http://journalarticle.ukm.my/7304/ Detection of shiga toxin 1 and 2 (stx1 and stx2) genes in escherichia coli 0157: H7 isolated from retail beef in Malaysia by multiplex polymerase chain reaction (PCR) A.M. Sahilah, H. Nor' Aishah, A. Ahmad Azuhairi, 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. Universiti Kebangsaan Malaysia 2010 Article PeerReviewed application/pdf en http://journalarticle.ukm.my/7304/1/03.pdf A.M. Sahilah, and H. Nor' Aishah, and A. Ahmad Azuhairi, (2010) Detection of shiga toxin 1 and 2 (stx1 and stx2) genes in escherichia coli 0157: H7 isolated from retail beef in Malaysia by multiplex polymerase chain reaction (PCR). Sains Malaysiana, 39 (1). pp. 57-63. ISSN 0126-6039 http://www.ukm.my/jsm/english_journals/vol39num1_2010/contentsVol39num1_2010.html
repository_type	Digital Repository
institution_category	Local University
institution	Universiti Kebangasaan Malaysia
building	UKM Institutional Repository
collection	Online Access
language	English
description	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.
format	Article
author	A.M. Sahilah, H. Nor' Aishah, A. Ahmad Azuhairi,
spellingShingle	A.M. Sahilah, H. Nor' Aishah, A. Ahmad Azuhairi, Detection of shiga toxin 1 and 2 (stx1 and stx2) genes in escherichia coli 0157: H7 isolated from retail beef in Malaysia by multiplex polymerase chain reaction (PCR)
author_facet	A.M. Sahilah, H. Nor' Aishah, A. Ahmad Azuhairi,
author_sort	A.M. Sahilah,
title	Detection of shiga toxin 1 and 2 (stx1 and stx2) genes in escherichia coli 0157: H7 isolated from retail beef in Malaysia by multiplex polymerase chain reaction (PCR)
title_short	Detection of shiga toxin 1 and 2 (stx1 and stx2) genes in escherichia coli 0157: H7 isolated from retail beef in Malaysia by multiplex polymerase chain reaction (PCR)
title_full	Detection of shiga toxin 1 and 2 (stx1 and stx2) genes in escherichia coli 0157: H7 isolated from retail beef in Malaysia by multiplex polymerase chain reaction (PCR)
title_fullStr	Detection of shiga toxin 1 and 2 (stx1 and stx2) genes in escherichia coli 0157: H7 isolated from retail beef in Malaysia by multiplex polymerase chain reaction (PCR)
title_full_unstemmed	Detection of shiga toxin 1 and 2 (stx1 and stx2) genes in escherichia coli 0157: H7 isolated from retail beef in Malaysia by multiplex polymerase chain reaction (PCR)
title_sort	detection of shiga toxin 1 and 2 (stx1 and stx2) genes in escherichia coli 0157: h7 isolated from retail beef in malaysia by multiplex polymerase chain reaction (pcr)
publisher	Universiti Kebangsaan Malaysia
publishDate	2010
url	http://journalarticle.ukm.my/7304/ http://journalarticle.ukm.my/7304/ http://journalarticle.ukm.my/7304/1/03.pdf
first_indexed	2023-09-18T19:49:20Z
last_indexed	2023-09-18T19:49:20Z
_version_	1777406113268891648

Detection of shiga toxin 1 and 2 (stx1 and stx2) genes in escherichia coli 0157: H7 isolated from retail beef in Malaysia by multiplex polymerase chain reaction (PCR)

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