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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| Format: | Thesis |
| Language: | English |
| Published: |
2010
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| Online Access: | http://eprints.usm.my/32387/ http://eprints.usm.my/32387/1/HASSANAIN_I._AL-TALIB_24%28NN%29.pdf |
| _version_ | 1848876798641700864 |
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| author | Al-Talib, Hassanain I. |
| author_facet | Al-Talib, Hassanain I. |
| author_sort | Al-Talib, Hassanain I. |
| building | USM Institutional Repository |
| collection | Online Access |
| 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. |
| first_indexed | 2025-11-15T17:05:17Z |
| format | Thesis |
| id | usm-32387 |
| institution | Universiti Sains Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T17:05:17Z |
| publishDate | 2010 |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | usm-323872017-05-15T04:43:29Z http://eprints.usm.my/32387/ Development Of A Thermostabilized Multiplex PCR Assay For The Rapid Detection Of Methicillin-Resistant Staphylococcus Aureus 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 NonPeerReviewed application/pdf en http://eprints.usm.my/32387/1/HASSANAIN_I._AL-TALIB_24%28NN%29.pdf Al-Talib, Hassanain I. (2010) Development Of A Thermostabilized Multiplex PCR Assay For The Rapid Detection Of Methicillin-Resistant Staphylococcus Aureus. PhD thesis, Universiti Sains Malaysia. |
| 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 |
| title | 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_short | 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 |
| topic | R5-130.5 General works |
| url | http://eprints.usm.my/32387/ http://eprints.usm.my/32387/1/HASSANAIN_I._AL-TALIB_24%28NN%29.pdf |