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

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.

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Abstract

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.

Item Type:	Thesis (PhD)
Subjects:	R Medicine > R Medicine (General) > R5-130.5 General works
Divisions:	Kampus Kesihatan (Health Campus) > Pusat Pengajian Sains Perubatan (School of Medical Sciences) > Thesis
Depositing User:	Mr Noorazilan Noordin
Date Deposited:	09 Mar 2017 08:42
Last Modified:	15 May 2017 04:43
URI:	http://eprints.usm.my/id/eprint/32387

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