Performance of MRSA screening at the Regensburg institute for medical microbiology and hygiene: an assessment of the value of broth enrichment
Gespeichert in:
| 1. Verfasser: | |
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| Format: | Abschlussarbeit Buch |
| Sprache: | English |
| Veröffentlicht: |
2008
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| Schlagworte: | |
| Online-Zugang: | Inhaltsverzeichnis |
| Beschreibung: | IV, 112 Bl. Ill., graph. Darst. |
Internformat
MARC
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| 245 | 1 | 0 | |a Performance of MRSA screening at the Regensburg institute for medical microbiology and hygiene |b an assessment of the value of broth enrichment |c von Welmoed Silvis |
| 264 | 1 | |c 2008 | |
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| 336 | |b txt |2 rdacontent | ||
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| 502 | |a Regensburg, Univ., Diss., 2008 | ||
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Datensatz im Suchindex
| _version_ | 1804138511573450752 |
|---|---|
| adam_text | Table of Contents
Contents
Page
1. Introduction 1
1.1. Object of study 3
2. Background 5
2.1. Staphylococcus aureus 5
2.1.1. Clinical relevance 5
2.1.2. Descriptive microbiology and identification 5
2.1.2.1. Colonial morphology 6
2.1.2.2. Catalase production 6
2.1.2.3. Coagulase production 6
2.1.3. P-lactam antibiotics: Mechanism of action 7
2.2. MRSA 9
2.2.1. Heterogeneous resistance 10
2.2.2. Inducing expression of resistance 10
2.3. Non-mecA-mediated oxacillin resistance 11
2.4. MRSA detection methods 12
2.4.1. Selective and differential media for MRSA screening 12
2.4.2. Broth enrichment 13
3. Materials and Methods 15
3.1. Subculture quantification experiments 15
3.1.1. Scale calibration 15
3.1.2. Swab absorption volume 15
3.1.3. Inoculation volume 16
3.1.4. Colony counts: Swab versus pipetted subculture 16
3.2. Experimental enrichment broths 17
3.2.1. Antimicrobial stock solutions 17
3.2.1.1. Quality control of antimicrobial stock solutions 17
3.2.1.2. Principles of broth microdilution 17
3.2.2. Preparation of broths 18
3.2.3. Quality control and shelf life of broths 19
3.2.4. Validation of broths 20
3.2.4.1. Selection of validation MRSA strains 20
3.2.4.2. Validation protocol 20
3.3. Pilot-study 22
3.4. Samples for MRSA screening 23
3.4.1. Patient group definition 23
3.4.2. Enrolment of samples 23
3.5. Workflow 25
3.5.1. Specimen processing 25
3.5.2. Current MRSA screening procedure (CSM) 25
3.5.2.1. Identification of S. aureus 27
3.5.2.2. Detection of methicillin resistance 27
3.5.3. Processing of enrichment broths 28
3.5.3.1. Identification of S. aumus and MRSA 29
3.6. Collected clinical MRSA isolates 31
3.6.1. Determination of MICs 31
II
Table of Contents
3.6.2. Molecular typing 31
3.6.3. Storage 31
3.7. Molecular techniques 32
3.7.1. DNA-isolation 32
3.7.2. Spa-typing 32
3.7.3. MLST 32
3.8. Statistical analysis 33
3.8.1. Statistical tests 33
3.8.2. Sample size 33
3.8.3. Gold standard 33
4. Results 34
4.1. Broth subculture procedure: a quantification 34
4.1.1. Scale calibration 34
4.1.2. Swab absorption volume 34
4.1.3. Swab inoculation volume 34
4.1.4. Swab-based versus pipette-based subculture 35
4.2. Broth validation 36
4.2.1. Validation strains 36
4.2.2. Broth validation results 38
4.2.2.1. Broths without antimicrobials: NAand TSB 38
4.2.2.2. Antimicrobial-containing broths 41
4.2.2.3. Growth support of PVL310 41
4.2.2.4. Positive subcultures of clear broths 42
4.2.2.5. Repeatability of results 42
4.2.3. Summary broth validation 43
4.3. Results pilot-study 45
4.4. Data characteristics 46
4.4.1. Samples included for study 46
4.4.2. Sample characteristics 46
4.5. Performance of CSM in MRSA-detection 49
4.5.1. Performance and sampling site 49
4.5.2. False-negative samples 51
4.5.3. MRSA-posltive samples exclusively detected by CSM 52
4.6. Effectiveness of broth enrichment 56
4.6.1. Broth performance 56
4.6.2. Effect of prolonged enrichment 58
4.6.3. Selectivity for MRSA 58
4.6.4. Performance of CSM plus one enrichment broth 58
4.6.5. Performance of two broths in parallel 59
4.6.6. Maximum MRSA yield 59
4.6.7. Broth performance and sampling site 59
4.6.8. Work-up time of broths 60
4.6.8.1. Broth turbidity 61
4.6.8.2. MRSA-quantfty in subculture 61
4.6.8.3. Gram-negative rods in subculture 61
4.7. Subculture on BBL™ CHROMagar™ MRSA 64
4.8. Collected clinical MRSA-Strains 66
4.8.1. Cefoxitin disk diffusion 66
4.8.2. MICs for ceftizoxime, cefoxitin and oxacillin 66
4.8.3. Spa-typing 67
III
Table of Contents
5. Discussion 70
5.1. Overview of major results 70
5.2. Generaiizibility of results 71
5.2.1. Collected MRSA isolates 72
5.3. Performance of CSM 73
5.4. Value of broth enrichment 75
5.4.1. HE 76
5.4.2. Poor performance of OX 76
5.4.3. The best broths: NA and CEF 77
5.4.4. No benefit of selective subculture on BD 78
5.4.5. Time to result and sensitivity 80
5.4.5.1. Visual detection of turbidity 81
5.4.6. Detection of MRSA spa-type t310 81
5.5. Gold standard MRSA yield 82
5.5.1. Cefoxitin inhibition zone diameter 85
5.6. Recommendations 88
6. Referencelist 91
7. Summary/Zusammenfassung 100
7.1. Summary 100
7.2. Zusammenfassung 102
8. List of abbreviations 104
9. Index of figures and tables 105
10. Appendix 106
11. Acknowledgements 109
12. Scientificoutput 110
13. Curriculumvitae 111
IV
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| dewey-ones | 614 - Forensic medicine; incidence of disease |
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| dewey-search | 614.44 |
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| discipline | Medizin |
| format | Thesis Book |
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| indexdate | 2024-07-09T21:29:21Z |
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| spelling | Silvis, Welmoed 1975- Verfasser (DE-588)137354673 aut Performance of MRSA screening at the Regensburg institute for medical microbiology and hygiene an assessment of the value of broth enrichment von Welmoed Silvis 2008 IV, 112 Bl. Ill., graph. Darst. txt rdacontent n rdamedia nc rdacarrier Regensburg, Univ., Diss., 2008 MRSA (DE-588)7501248-0 gnd rswk-swf Screening (DE-588)4054045-5 gnd rswk-swf (DE-588)4113937-9 Hochschulschrift gnd-content MRSA (DE-588)7501248-0 s Screening (DE-588)4054045-5 s b DE-604 HBZ Datenaustausch application/pdf http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=017045631&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA Inhaltsverzeichnis |
| spellingShingle | Silvis, Welmoed 1975- Performance of MRSA screening at the Regensburg institute for medical microbiology and hygiene an assessment of the value of broth enrichment MRSA (DE-588)7501248-0 gnd Screening (DE-588)4054045-5 gnd |
| subject_GND | (DE-588)7501248-0 (DE-588)4054045-5 (DE-588)4113937-9 |
| title | Performance of MRSA screening at the Regensburg institute for medical microbiology and hygiene an assessment of the value of broth enrichment |
| title_auth | Performance of MRSA screening at the Regensburg institute for medical microbiology and hygiene an assessment of the value of broth enrichment |
| title_exact_search | Performance of MRSA screening at the Regensburg institute for medical microbiology and hygiene an assessment of the value of broth enrichment |
| title_full | Performance of MRSA screening at the Regensburg institute for medical microbiology and hygiene an assessment of the value of broth enrichment von Welmoed Silvis |
| title_fullStr | Performance of MRSA screening at the Regensburg institute for medical microbiology and hygiene an assessment of the value of broth enrichment von Welmoed Silvis |
| title_full_unstemmed | Performance of MRSA screening at the Regensburg institute for medical microbiology and hygiene an assessment of the value of broth enrichment von Welmoed Silvis |
| title_short | Performance of MRSA screening at the Regensburg institute for medical microbiology and hygiene |
| title_sort | performance of mrsa screening at the regensburg institute for medical microbiology and hygiene an assessment of the value of broth enrichment |
| title_sub | an assessment of the value of broth enrichment |
| topic | MRSA (DE-588)7501248-0 gnd Screening (DE-588)4054045-5 gnd |
| topic_facet | MRSA Screening Hochschulschrift |
| url | http://bvbr.bib-bvb.de:8991/F?func=service&doc_library=BVB01&local_base=BVB01&doc_number=017045631&sequence=000002&line_number=0001&func_code=DB_RECORDS&service_type=MEDIA |
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