Molecular Characterisation Of Escherichia Coli Serogroup O 157:H7

Fourteen E. coli O 157:H7 beef isolates were characterised by using four specific epidemiological markers: combination of antibiogram and plasmid profiling, pulsed-field gel electrophoresis (PFGE) and arbitrary primed polymerase chain reaction (AP-PCR). These markers were assessed for their reli...

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Main Author:	Mohd Dzomir, Ahmad Zainuri
Format:	Thesis
Language:	English
Published:	1998
Subjects:	Escherichia coli
Online Access:	http://psasir.upm.edu.my/id/eprint/8384/1/FSMB_1998_1_A.pdf
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id	my-upm-ir.8384
record_format	uketd_dc
spelling	my-upm-ir.83842016-07-22T03:18:29Z Molecular Characterisation Of Escherichia Coli Serogroup O 157:H7 1998 Mohd Dzomir, Ahmad Zainuri Fourteen E. coli O 157:H7 beef isolates were characterised by using four specific epidemiological markers: combination of antibiogram and plasmid profiling, pulsed-field gel electrophoresis (PFGE) and arbitrary primed polymerase chain reaction (AP-PCR). These markers were assessed for their reliability, typability, rapidity and discriminatory power in differentiating beef E. coli O 157:H7 strains from different locations, namely Bangsar, Kajang, Petaling Jaya and Serdang. The majority of the isolates were resistant to penicillin G (100%), vancomycin (100%), trimethoprim/ sulphametoxazole (100%), bacitracin ( 1 00%) and erythromycin (92.8%). Only 21.4% were resistant to carbenicillin. 14.3% and 7.1% were resistant to ampicillin and cephalotin, respectively Plasmid analysis revealed three basic plasmid patterns among E. coli O 157:H7 strains, profile 1 characterised by plasmid DNA of 60 and 2.5 MDa, profile 2 characterised by plasmid of 60 MDa, and profile 3 characterised by the absence of any plasmid in the strains. Grouping according to combination of antibiogram and plasmid analysis indicated eight different groups as two strains with similar antibiotype could be distinguished into two different strains by their dissimilar plasmid profile. However, the reliability of antibiogram and plasmid analysis in typing E. coli O 157:H7 can be questioned. Thus, other reliable methods such as PFGE and AP-PCR were then applied. In the present study, macrorestriction of genomic DNA of E. coli 0 1 57:H7 using Xbal, SpeI and HindIII and analysed by PFGE successfully grouped ten out of fourteen isolates into five groups and provided evidence of epidemiologically related strains between strains of different and same locations. However, AP-PCR using three short primers grouped the isolates into fourteen distinct groups and differentiates isolates that were not differentiated by PFGE. The overall analysis of the present study revealed AP-PCR as the most suitable method to differentiate E. coli O 157:H7 because it was more discriminatory, less labor intensive and applicable to all isolates. Using this method, it was clearly shown that all fourteen E. coli O 157:H7 existed as independent isolates. Escherichia coli 1998 Thesis http://psasir.upm.edu.my/id/eprint/8384/ http://psasir.upm.edu.my/id/eprint/8384/1/FSMB_1998_1_A.pdf application/pdf en public masters Universiti Putra Malaysia Escherichia coli Food Science and Technology
institution	Universiti Putra Malaysia
collection	PSAS Institutional Repository
language	English
topic	Escherichia coli
spellingShingle	Escherichia coli Mohd Dzomir, Ahmad Zainuri Molecular Characterisation Of Escherichia Coli Serogroup O 157:H7
description	Fourteen E. coli O 157:H7 beef isolates were characterised by using four specific epidemiological markers: combination of antibiogram and plasmid profiling, pulsed-field gel electrophoresis (PFGE) and arbitrary primed polymerase chain reaction (AP-PCR). These markers were assessed for their reliability, typability, rapidity and discriminatory power in differentiating beef E. coli O 157:H7 strains from different locations, namely Bangsar, Kajang, Petaling Jaya and Serdang. The majority of the isolates were resistant to penicillin G (100%), vancomycin (100%), trimethoprim/ sulphametoxazole (100%), bacitracin ( 1 00%) and erythromycin (92.8%). Only 21.4% were resistant to carbenicillin. 14.3% and 7.1% were resistant to ampicillin and cephalotin, respectively Plasmid analysis revealed three basic plasmid patterns among E. coli O 157:H7 strains, profile 1 characterised by plasmid DNA of 60 and 2.5 MDa, profile 2 characterised by plasmid of 60 MDa, and profile 3 characterised by the absence of any plasmid in the strains. Grouping according to combination of antibiogram and plasmid analysis indicated eight different groups as two strains with similar antibiotype could be distinguished into two different strains by their dissimilar plasmid profile. However, the reliability of antibiogram and plasmid analysis in typing E. coli O 157:H7 can be questioned. Thus, other reliable methods such as PFGE and AP-PCR were then applied. In the present study, macrorestriction of genomic DNA of E. coli 0 1 57:H7 using Xbal, SpeI and HindIII and analysed by PFGE successfully grouped ten out of fourteen isolates into five groups and provided evidence of epidemiologically related strains between strains of different and same locations. However, AP-PCR using three short primers grouped the isolates into fourteen distinct groups and differentiates isolates that were not differentiated by PFGE. The overall analysis of the present study revealed AP-PCR as the most suitable method to differentiate E. coli O 157:H7 because it was more discriminatory, less labor intensive and applicable to all isolates. Using this method, it was clearly shown that all fourteen E. coli O 157:H7 existed as independent isolates.
format	Thesis
qualification_level	Master's degree
author	Mohd Dzomir, Ahmad Zainuri
author_facet	Mohd Dzomir, Ahmad Zainuri
author_sort	Mohd Dzomir, Ahmad Zainuri
title	Molecular Characterisation Of Escherichia Coli Serogroup O 157:H7
title_short	Molecular Characterisation Of Escherichia Coli Serogroup O 157:H7
title_full	Molecular Characterisation Of Escherichia Coli Serogroup O 157:H7
title_fullStr	Molecular Characterisation Of Escherichia Coli Serogroup O 157:H7
title_full_unstemmed	Molecular Characterisation Of Escherichia Coli Serogroup O 157:H7
title_sort	molecular characterisation of escherichia coli serogroup o 157:h7
granting_institution	Universiti Putra Malaysia
granting_department	Food Science and Technology
publishDate	1998
url	http://psasir.upm.edu.my/id/eprint/8384/1/FSMB_1998_1_A.pdf
_version_	1747810793946087424

Molecular Characterisation Of Escherichia Coli Serogroup O 157:H7

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