For the first time, an in vitro cystic fibrosis (CF) artificial sputum model (ASM) was found to support the growth and survival of a clinical epidemic strain of meticillin-resistant Staphylococcus aureus (EMRSAl6-252). Specific components, which included mucin. DNA and others, were removed from ASM and the physiological impact of this was fully explored using viable counts and light microscopy. As CF patients are known to develop cystic fibrosis-related diabetes or CFRD, glucose was added to ASM (GASM), to explore the physiological impact of glucose on the growth and survival MRSA252. Total RNA was extracted from the corresponding log phases of MRSA252 grown in brain heart infusion (BHI) as a laboratory control, as well as ASM and GASM. RNA was extracted in order to conduct microarray analysis. MRSA252 DNA was used as a control. RNA (from the samples) was labelled with Cy5 and control DNA was labelled with Cy3. Once labelled and amplified, the Cy5/Cy3 mixture was then purified and hybridised onto an array containing seven sequenced S. aureus : genomes (N315, Mu50, MW2, MRSA252, MSSA476, COL and NCTC8325).
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:501769 |
| Date | January 2008 |
| Creators | Micallef, Christianne |
| Publisher | Kingston University |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | http://eprints.kingston.ac.uk/20405/ |
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