Quantitative PCR analysis of genes expressed during biofilm development of methicillin resistant Staphylococcus aureus (MRSA)
Staphylococcus aureus biofilm associated infections remains a major clinical concern in patients with indwelling devices. Quantitative real-time PCR (qPCR) can be used to investigate the pathogenic role of such biofilms. We describe qPCRs for 12 adhesion and biofilm-related genes of four S. aureus i...
| Main Authors: | , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Published: |
Elsevier
2013
|
| _version_ | 1796971559873675264 |
|---|---|
| author | Atshan, Salman Sahab Shamsudin, Mariana Nor Karunanidhi, Arunkumar Belkum, Alex van Lung, Leslie Than Thian Sekawi, Zamberi Nathan, Jayakayatri Jeevajothi Ling, King Hwa Seng, Johnson Shueh Chong Alreshidi, Mateq Ali Abduljaleel, Salwa A. Awang Hamat, Rukman |
| author_facet | Atshan, Salman Sahab Shamsudin, Mariana Nor Karunanidhi, Arunkumar Belkum, Alex van Lung, Leslie Than Thian Sekawi, Zamberi Nathan, Jayakayatri Jeevajothi Ling, King Hwa Seng, Johnson Shueh Chong Alreshidi, Mateq Ali Abduljaleel, Salwa A. Awang Hamat, Rukman |
| author_sort | Atshan, Salman Sahab |
| collection | UPM |
| description | Staphylococcus aureus biofilm associated infections remains a major clinical concern in patients with indwelling devices. Quantitative real-time PCR (qPCR) can be used to investigate the pathogenic role of such biofilms. We describe qPCRs for 12 adhesion and biofilm-related genes of four S. aureus isolates which were applied during in vitro biofilm development. An endogenous control (16S rRNA) was used for signal normalization. We compared the qPCR results with structural analysis using scanning electron microscopy (SEM). The SEM studies showed different cellular products surrounding the aggregated cells at different times of biofilm formation. Using qPCR, we found that expression levels of the gene encoding fibronectin binding protein A and B and clumping factor B (fnbA/B and clfB), which involves in primary adherence of S. aureus, were significantly increased at 24h and decreased slightly and variably at 48 h when all 4 isolates were considered. The elastin binding protein (ebps) RNA expression level was significantly enhanced more than 6-fold at 24 and 48 h compared to 12h. Similar results were obtained for the intercellular adhesion biofilm required genes type C (icaC). In addition, qPCR revealed a fluctuation in expression levels at different time points of biofilm growth of other genes, indicating that different parameter modes of growth processes are operating at different times. |
| first_indexed | 2024-03-06T08:14:48Z |
| format | Article |
| id | upm.eprints-29540 |
| institution | Universiti Putra Malaysia |
| last_indexed | 2024-03-06T08:14:48Z |
| publishDate | 2013 |
| publisher | Elsevier |
| record_format | dspace |
| spelling | upm.eprints-295402015-12-08T03:56:51Z http://psasir.upm.edu.my/id/eprint/29540/ Quantitative PCR analysis of genes expressed during biofilm development of methicillin resistant Staphylococcus aureus (MRSA) Atshan, Salman Sahab Shamsudin, Mariana Nor Karunanidhi, Arunkumar Belkum, Alex van Lung, Leslie Than Thian Sekawi, Zamberi Nathan, Jayakayatri Jeevajothi Ling, King Hwa Seng, Johnson Shueh Chong Alreshidi, Mateq Ali Abduljaleel, Salwa A. Awang Hamat, Rukman Staphylococcus aureus biofilm associated infections remains a major clinical concern in patients with indwelling devices. Quantitative real-time PCR (qPCR) can be used to investigate the pathogenic role of such biofilms. We describe qPCRs for 12 adhesion and biofilm-related genes of four S. aureus isolates which were applied during in vitro biofilm development. An endogenous control (16S rRNA) was used for signal normalization. We compared the qPCR results with structural analysis using scanning electron microscopy (SEM). The SEM studies showed different cellular products surrounding the aggregated cells at different times of biofilm formation. Using qPCR, we found that expression levels of the gene encoding fibronectin binding protein A and B and clumping factor B (fnbA/B and clfB), which involves in primary adherence of S. aureus, were significantly increased at 24h and decreased slightly and variably at 48 h when all 4 isolates were considered. The elastin binding protein (ebps) RNA expression level was significantly enhanced more than 6-fold at 24 and 48 h compared to 12h. Similar results were obtained for the intercellular adhesion biofilm required genes type C (icaC). In addition, qPCR revealed a fluctuation in expression levels at different time points of biofilm growth of other genes, indicating that different parameter modes of growth processes are operating at different times. Elsevier 2013-08 Article PeerReviewed Atshan, Salman Sahab and Shamsudin, Mariana Nor and Karunanidhi, Arunkumar and Belkum, Alex van and Lung, Leslie Than Thian and Sekawi, Zamberi and Nathan, Jayakayatri Jeevajothi and Ling, King Hwa and Seng, Johnson Shueh Chong and Alreshidi, Mateq Ali and Abduljaleel, Salwa A. and Awang Hamat, Rukman (2013) Quantitative PCR analysis of genes expressed during biofilm development of methicillin resistant Staphylococcus aureus (MRSA). Infection, Genetics and Evolution, 18. pp. 106-112. ISSN 1567-1348; ESSN: 1567-7257 10.1016/j.meegid.2013.05.002 |
| spellingShingle | Atshan, Salman Sahab Shamsudin, Mariana Nor Karunanidhi, Arunkumar Belkum, Alex van Lung, Leslie Than Thian Sekawi, Zamberi Nathan, Jayakayatri Jeevajothi Ling, King Hwa Seng, Johnson Shueh Chong Alreshidi, Mateq Ali Abduljaleel, Salwa A. Awang Hamat, Rukman Quantitative PCR analysis of genes expressed during biofilm development of methicillin resistant Staphylococcus aureus (MRSA) |
| title | Quantitative PCR analysis of genes expressed during biofilm development of methicillin resistant Staphylococcus aureus (MRSA) |
| title_full | Quantitative PCR analysis of genes expressed during biofilm development of methicillin resistant Staphylococcus aureus (MRSA) |
| title_fullStr | Quantitative PCR analysis of genes expressed during biofilm development of methicillin resistant Staphylococcus aureus (MRSA) |
| title_full_unstemmed | Quantitative PCR analysis of genes expressed during biofilm development of methicillin resistant Staphylococcus aureus (MRSA) |
| title_short | Quantitative PCR analysis of genes expressed during biofilm development of methicillin resistant Staphylococcus aureus (MRSA) |
| title_sort | quantitative pcr analysis of genes expressed during biofilm development of methicillin resistant staphylococcus aureus mrsa |
| work_keys_str_mv | AT atshansalmansahab quantitativepcranalysisofgenesexpressedduringbiofilmdevelopmentofmethicillinresistantstaphylococcusaureusmrsa AT shamsudinmariananor quantitativepcranalysisofgenesexpressedduringbiofilmdevelopmentofmethicillinresistantstaphylococcusaureusmrsa AT karunanidhiarunkumar quantitativepcranalysisofgenesexpressedduringbiofilmdevelopmentofmethicillinresistantstaphylococcusaureusmrsa AT belkumalexvan quantitativepcranalysisofgenesexpressedduringbiofilmdevelopmentofmethicillinresistantstaphylococcusaureusmrsa AT lunglesliethanthian quantitativepcranalysisofgenesexpressedduringbiofilmdevelopmentofmethicillinresistantstaphylococcusaureusmrsa AT sekawizamberi quantitativepcranalysisofgenesexpressedduringbiofilmdevelopmentofmethicillinresistantstaphylococcusaureusmrsa AT nathanjayakayatrijeevajothi quantitativepcranalysisofgenesexpressedduringbiofilmdevelopmentofmethicillinresistantstaphylococcusaureusmrsa AT lingkinghwa quantitativepcranalysisofgenesexpressedduringbiofilmdevelopmentofmethicillinresistantstaphylococcusaureusmrsa AT sengjohnsonshuehchong quantitativepcranalysisofgenesexpressedduringbiofilmdevelopmentofmethicillinresistantstaphylococcusaureusmrsa AT alreshidimateqali quantitativepcranalysisofgenesexpressedduringbiofilmdevelopmentofmethicillinresistantstaphylococcusaureusmrsa AT abduljaleelsalwaa quantitativepcranalysisofgenesexpressedduringbiofilmdevelopmentofmethicillinresistantstaphylococcusaureusmrsa AT awanghamatrukman quantitativepcranalysisofgenesexpressedduringbiofilmdevelopmentofmethicillinresistantstaphylococcusaureusmrsa |