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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| Format: | Article |
| Language: | English |
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Universiti Kebangsaan Malaysia
2010
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| Online Access: | http://journalarticle.ukm.my/7304/1/03.pdf |
| _version_ | 1796930514450382848 |
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| author | A.M. Sahilah, H. Nor' Aishah, A. Ahmad Azuhairi, |
| author_facet | A.M. Sahilah, H. Nor' Aishah, A. Ahmad Azuhairi, |
| author_sort | A.M. Sahilah, |
| collection | UKM |
| 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. |
| first_indexed | 2024-03-06T04:04:10Z |
| format | Article |
| id | ukm.eprints-7304 |
| institution | Universiti Kebangsaan Malaysia |
| language | English |
| last_indexed | 2024-03-06T04:04:10Z |
| publishDate | 2010 |
| publisher | Universiti Kebangsaan Malaysia |
| record_format | dspace |
| spelling | ukm.eprints-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 |
| 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) |
| 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_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_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_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 |
| url | http://journalarticle.ukm.my/7304/1/03.pdf |
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