Development Of A Thermostabilized Multiplex PCR Assay For The Rapid Detection Of Methicillin-Resistant Staphylococcus Aureus

Staphylococcus aureus rintang methicillin (MRSA) bertanggungjawab terhadap kebanyakan jangkitan nosokomial dan komuniti. Ujian kultur konvensional mengambil masa selama dua hingga lima hari untuk menghasilkan maklumat penuh mengenai organisma dan pola kerintangan antibiotiknya. Oleh itu, kajian i...

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Main Author:	Al-Talib, Hassanain I.
Format:	Thesis
Language:	English
Published:	2010
Subjects:	R5-130.5 General works
Online Access:	http://eprints.usm.my/32387/1/HASSANAIN_I._AL-TALIB_24%28NN%29.pdf
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id	my-usm-ep.32387
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spelling	my-usm-ep.323872017-05-15T04:43:29Z Development Of A Thermostabilized Multiplex PCR Assay For The Rapid Detection Of Methicillin-Resistant Staphylococcus Aureus 2010-03 Al-Talib, Hassanain I. R5-130.5 General works Staphylococcus aureus rintang methicillin (MRSA) bertanggungjawab terhadap kebanyakan jangkitan nosokomial dan komuniti. Ujian kultur konvensional mengambil masa selama dua hingga lima hari untuk menghasilkan maklumat penuh mengenai organisma dan pola kerintangan antibiotiknya. Oleh itu, kajian ini bertujuan untuk membangunkan ujian reaksi berantai polimerasi berganda untuk pengesanan MRSA dengan pantas. Ujian ini akan mengesan lima gen iaitu 16S rRNA gen dari genus Staphylococcus,femA Staphylococcus aureus, mecA yang mengekod rintangan methicillin, lukS yang mengekod pengeluaran leukosidin Panton-Valentine (PVL), sitotoksin nekrosis, dan satu gen kawalan dalaman secara serentak. Pasangan primer yang unik dan khusus telah pireka untuk mengamplifikasi lima gen dengan produk reaksi berantai polimerasi pada julat 151 hingga 759 bp. Primer yang spesifik disahkan berdasarkan urutan jujukan DNA produk reaksi berantai polimerasi berganda dan analisa Blast. The emergence of methicillin-resistant Staphylococcus aureus (MRSA) is responsible for nosocomial and community-acquired infections. The conventional culture test takes 2-5 days to yield complete information of the organism and its antibiotic sensitivity pattern. Hence our present study was focused on developing a multiplex PCR assay for the rapid detection of MRSA. The assay simultaneously detected five genes, namely 16S rRNA of the Staphylococcus genus,femA of S. aureus, mecA that encodes methicillin resistance, lukS that encodes production of Panton- Valentine leukocidin (PVL), a necrotizing cytotoxin and one internal control. Unique and specific primer pairs were designed to amplify the 5 genes with the PCR products ranging from 151 to 759 bp. The specificity of the primers was confirmed by DNA sequencing of the multiplex PCR products and BLAST analysis. 2010-03 Thesis http://eprints.usm.my/32387/ http://eprints.usm.my/32387/1/HASSANAIN_I._AL-TALIB_24%28NN%29.pdf application/pdf en public phd doctoral Universiti Sains Malaysia Pusat Pengajian Sains Perubatan (School of Medical Sciences)
institution	Universiti Sains Malaysia
collection	USM Institutional Repository
language	English
topic	R5-130.5 General works
spellingShingle	R5-130.5 General works Al-Talib, Hassanain I. Development Of A Thermostabilized Multiplex PCR Assay For The Rapid Detection Of Methicillin-Resistant Staphylococcus Aureus
description	Staphylococcus aureus rintang methicillin (MRSA) bertanggungjawab terhadap kebanyakan jangkitan nosokomial dan komuniti. Ujian kultur konvensional mengambil masa selama dua hingga lima hari untuk menghasilkan maklumat penuh mengenai organisma dan pola kerintangan antibiotiknya. Oleh itu, kajian ini bertujuan untuk membangunkan ujian reaksi berantai polimerasi berganda untuk pengesanan MRSA dengan pantas. Ujian ini akan mengesan lima gen iaitu 16S rRNA gen dari genus Staphylococcus,femA Staphylococcus aureus, mecA yang mengekod rintangan methicillin, lukS yang mengekod pengeluaran leukosidin Panton-Valentine (PVL), sitotoksin nekrosis, dan satu gen kawalan dalaman secara serentak. Pasangan primer yang unik dan khusus telah pireka untuk mengamplifikasi lima gen dengan produk reaksi berantai polimerasi pada julat 151 hingga 759 bp. Primer yang spesifik disahkan berdasarkan urutan jujukan DNA produk reaksi berantai polimerasi berganda dan analisa Blast. The emergence of methicillin-resistant Staphylococcus aureus (MRSA) is responsible for nosocomial and community-acquired infections. The conventional culture test takes 2-5 days to yield complete information of the organism and its antibiotic sensitivity pattern. Hence our present study was focused on developing a multiplex PCR assay for the rapid detection of MRSA. The assay simultaneously detected five genes, namely 16S rRNA of the Staphylococcus genus,femA of S. aureus, mecA that encodes methicillin resistance, lukS that encodes production of Panton- Valentine leukocidin (PVL), a necrotizing cytotoxin and one internal control. Unique and specific primer pairs were designed to amplify the 5 genes with the PCR products ranging from 151 to 759 bp. The specificity of the primers was confirmed by DNA sequencing of the multiplex PCR products and BLAST analysis.
format	Thesis
qualification_name	Doctor of Philosophy (PhD.)
qualification_level	Doctorate
author	Al-Talib, Hassanain I.
author_facet	Al-Talib, Hassanain I.
author_sort	Al-Talib, Hassanain I.
title	Development Of A Thermostabilized Multiplex PCR Assay For The Rapid Detection Of Methicillin-Resistant Staphylococcus Aureus
title_short	Development Of A Thermostabilized Multiplex PCR Assay For The Rapid Detection Of Methicillin-Resistant Staphylococcus Aureus
title_full	Development Of A Thermostabilized Multiplex PCR Assay For The Rapid Detection Of Methicillin-Resistant Staphylococcus Aureus
title_fullStr	Development Of A Thermostabilized Multiplex PCR Assay For The Rapid Detection Of Methicillin-Resistant Staphylococcus Aureus
title_full_unstemmed	Development Of A Thermostabilized Multiplex PCR Assay For The Rapid Detection Of Methicillin-Resistant Staphylococcus Aureus
title_sort	development of a thermostabilized multiplex pcr assay for the rapid detection of methicillin-resistant staphylococcus aureus
granting_institution	Universiti Sains Malaysia
granting_department	Pusat Pengajian Sains Perubatan (School of Medical Sciences)
publishDate	2010
url	http://eprints.usm.my/32387/1/HASSANAIN_I._AL-TALIB_24%28NN%29.pdf
_version_	1747820572384952320

Development Of A Thermostabilized Multiplex PCR Assay For The Rapid Detection Of Methicillin-Resistant Staphylococcus Aureus

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