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Samenvatting Medische bacteriologie - MLT2 - Hogent

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Samenvatting van de powerpoint presentaties. Ik heb alle leerdoelen en het extra document met aandachtspunten overlopen. Enkel hoofdstuk 9 (moest niet gekend zijn) en hoofdstuk 13 heb ik niet samengevat. Verder heb ik pas achteraf de aandachtspunten overlopen dus bij hoofdstuk 4 staan er onderwerpe...

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  • June 8, 2023
  • June 8, 2023
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Medische bacteriologie
Inhoud
H0. Inleiding ...................................................................................................................................................................... 7
0.1. Pathogene micro-organismen ........................................................................................................................... 7
1. Terrein van de medische bacteriologie ............................................................................................................. 7
2. Pathogene µorganismen ................................................................................................................................... 7
3. Classificatie van µorganismen ........................................................................................................................... 7
0.2. Morfologie en kokken ....................................................................................................................................... 8
Morfologische classificatie ........................................................................................................................................ 8
Filamenteuze bacteriën............................................................................................................................................. 8
Echte bacteriën ......................................................................................................................................................... 8
Kokken ....................................................................................................................................................................... 8
Bacillen ...................................................................................................................................................................... 8
Vibrio’s (gram -) ......................................................................................................................................................... 8
Spirillen (gram -) ........................................................................................................................................................ 8
Spirocheten ............................................................................................................................................................... 8
0.3. Taxonomie ......................................................................................................................................................... 9
Biochemische classificatie ......................................................................................................................................... 9
Numerische classificatie ............................................................................................................................................ 9
Fylogenetische classificatie ....................................................................................................................................... 9
0.4. De celwand van de bacterie ............................................................................................................................ 10
H1. Pathogeniciteit .......................................................................................................................................................... 11
1.1. Principes van ziekte en epidemiologie ............................................................................................................ 11
Enkele begrippen .................................................................................................................................................... 11
Normale flora .......................................................................................................................................................... 11
Het optreden van een ziekte ................................................................................................................................... 12
Soorten infectieuze ziekten ..................................................................................................................................... 12
Betrokkenheid van de gastheer .............................................................................................................................. 13
Verspreiding van een ziekte .................................................................................................................................... 13
Nosocomiale infecties ............................................................................................................................................. 14
1.2. Microbiële mechanismen van pathogeniciteit................................................................................................ 14
Het binnendringen van een µorganisme in het lichaam: beïnvloedende (virulentie) factoren en mechanismen . 14
Hoe brengt een pathogeen schade aan bij de gastheer? ....................................................................................... 16
Plasmiden, lyosgeniciteit, pathogeniciteit .............................................................................................................. 17
H2. Microscopisch onderzoek ......................................................................................................................................... 18
2.1 Onderdeel van een microscoop ............................................................................................................................ 18
2.2 Soorten .................................................................................................................................................................. 18
2.3 Vers onderzoek...................................................................................................................................................... 18

1

, 2.4 De kleuringen ........................................................................................................................................................ 19
Gramkleuring........................................................................................................................................................... 19
De Ziehl-Neelsen-kleuring ....................................................................................................................................... 19
2.6 De auraminekleuring ......................................................................................................................................... 20
2.7 Immunofluorescentiekleuring ........................................................................................................................... 20
2.8 Flagellakleuring ................................................................................................................................................. 20
2.9 Giemsakleuring ................................................................................................................................................. 20
2.10 Calcofluorkleuring ........................................................................................................................................... 20
H3. Kweekmethoden ....................................................................................................................................................... 21
3.1 Algemeen .............................................................................................................................................................. 21
3.2 Kweekmethoden ................................................................................................................................................... 21
3.2.1 Vaste en vloeibare voedingsbodems.............................................................................................................. 21
3.2.2 Selectieve voedingsbodems ........................................................................................................................... 22
3.2.3 Aanrijkingsbodems ......................................................................................................................................... 24
3.2.4 Specifieke isolatiebodems .............................................................................................................................. 24
3.3 Entingstechnieken ................................................................................................................................................. 24
Het trekken van een reincultuur ............................................................................................................................. 24
3.4 Incubatie-omstandigheden ................................................................................................................................... 24
H4. Biochemische identificatiemethoden ....................................................................................................................... 25
4.1. Primaire kenmerken ............................................................................................................................................. 25
Morfologische kenmerken ...................................................................................................................................... 25
Groeivoorwaarden .................................................................................................................................................. 25
Gevoeligheid voor chemische stoffen en antibiotica .............................................................................................. 26
Pigmentproductie.................................................................................................................................................... 26
Hemolyse (!!)........................................................................................................................................................... 26
Snelle enzymatische testen ..................................................................................................................................... 27
Latexagglutinatietesten ........................................................................................................................................... 27
4.2. Secundaire kenmerken (enkel voor practicum open boek test) .......................................................................... 28
Afbraak van koolhydraten ....................................................................................................................................... 28
Afbraak van eiwitten, aminozuren en DNA ............................................................................................................. 30
Gebruik van eenvoudige C-bron.............................................................................................................................. 31
Urease test .............................................................................................................................................................. 32
Nitraatreductietest (niet) ........................................................................................................................................ 32
Afbraak van lipiden (niet) ........................................................................................................................................ 32
4.3. Gecombineerde multitestmedia .......................................................................................................................... 33
Kligler Iron Agar (KIA) .............................................................................................................................................. 33
Triple Sugar Iron (TSI) → lezen ............................................................................................................................... 33
Mobiliteit- Indol- Urease medium (MIU) ................................................................................................................ 34
Mobiliteit-Indol-Ornithine medium (MIO) → lezen ................................................................................................ 34

2

, Lysine Iron Agar (LIA)............................................................................................................................................... 34
4.4. Commerciële biochemische identificatiesystemen.............................................................................................. 35
API-20E..................................................................................................................................................................... 35
Chromogene media ................................................................................................................................................. 35
4.5. MALDI-TOF-MS ..................................................................................................................................................... 35
MALDI: Matrix Assisted Laser Desorption Ionization .............................................................................................. 35
TOF: Time of flight + MS: massaspectrum .............................................................................................................. 35
H5. Moleculaire technieken in de bacteriologie ............................................................................................................. 37
5.4. Moleculaire technieken in het microbiologisch lab ............................................................................................. 37
5.4.1. Aantonen aanwezigheid van een species in een klinisch staal ..................................................................... 37
5.4.2. Genotypering van stammen via fingerprinting ............................................................................................. 39
5.5. Huidige diagnostiek .............................................................................................................................................. 41
H6. Enterobacteriaceae................................................................................................................................................... 42
6.1. Gram-negatieve bacteriën ................................................................................................................................... 42
Groepen bij gram-negatieven ................................................................................................................................. 42
Aantal belangrijke en te kennen kenmerken .......................................................................................................... 42
6.2. Geslachten en algemene kenmerken ................................................................................................................... 42
Gemeenschappelijk kenmerken .............................................................................................................................. 42
Indeling op basis van pathogeen vermogen ........................................................................................................... 42
6.3. Cultuur en biochemische identificatie ................................................................................................................. 43
6.4. De Enteropathogenen .......................................................................................................................................... 43
6.3.1 Salmonella enterica ........................................................................................................................................ 43
6.3.2. Shigella spp. .................................................................................................................................................. 44
6.3.3. Yersinia Spp. .................................................................................................................................................. 44
6.3.4. E. coli ............................................................................................................................................................. 45
6.4. Opportunistische pathogenen: Enterobacter spp................................................................................................ 45
6.4.1. Enterobacter spp. .......................................................................................................................................... 45
6.4.2. Klebsiella spp................................................................................................................................................. 45
6.4.3. Proteus – Providencia – Morganella = Transiënte flora (UREASE POSITIEF) ................................................. 46
6.4.4. Serratia spp. .................................................................................................................................................. 46
6.4.5. Citrobacter spp. ............................................................................................................................................. 46
6.4.6. Hafnia alvei.................................................................................................................................................... 46
6.4.7. Edwarsiella tarda ........................................................................................................................................... 46
6.4.8. Andere Enterobacteriaceae .......................................................................................................................... 46
H7. Oxidase-positieve fermenterende gram-negatieven ............................................................................................... 47
7.1. Geslachten en algemene kenmerken ................................................................................................................... 47
7.2. Vibrio spp. ............................................................................................................................................................ 47
Specifieke kenmerken ............................................................................................................................................. 47
Pathogenese ............................................................................................................................................................ 47

3

, Pathogeen vermogen .............................................................................................................................................. 47
Staal ......................................................................................................................................................................... 47
Isolatie en identificatie ............................................................................................................................................ 48
7.3. Aeromonas en Plasiomonas spp. ......................................................................................................................... 48
Specifieke kenmerken ............................................................................................................................................. 48
Pathogenese ............................................................................................................................................................ 48
Pathogeen vermogen: na watercontact! ................................................................................................................ 48
Staal ......................................................................................................................................................................... 48
Isolatie en identificatie ............................................................................................................................................ 48
7.4. Pasteurella spp. .................................................................................................................................................... 49
Specifieke kenmerken ............................................................................................................................................. 49
Pathogenese ............................................................................................................................................................ 49
Pathogeen vermogen .............................................................................................................................................. 49
Staal ......................................................................................................................................................................... 49
Isolatie en identificatie ............................................................................................................................................ 49
H8. Niet-vergistende aërobe gram-negatieve bacillen KK H2S- ................................................................................ 50
8.1. Algemene eigenschappen en soorten.................................................................................................................. 50
8.2. Pseudomonas aeruginosa .................................................................................................................................... 50
Specifieke kenmerken ............................................................................................................................................. 50
Pathogenese ............................................................................................................................................................ 50
Pathogeen vermogen .............................................................................................................................................. 50
Staal ......................................................................................................................................................................... 50
Isolatie en identificatie ............................................................................................................................................ 50
8.3. Xanthomonas spp. ............................................................................................................................................... 51
Specifieke kenmerken ............................................................................................................................................. 51
Pathogenese ............................................................................................................................................................ 51
Pathogeen vermogen .............................................................................................................................................. 51
Staal ......................................................................................................................................................................... 51
Isolatie en identificatie ............................................................................................................................................ 51
8.4. Alcaligenes spp. .................................................................................................................................................... 51
Specifieke kenmerken ............................................................................................................................................. 51
Pathogenese ............................................................................................................................................................ 51
Pathogeen vermogen .............................................................................................................................................. 51
Staal ......................................................................................................................................................................... 51
Isolatie en identificatie ............................................................................................................................................ 51
8.5. Acinetobacter spp. ............................................................................................................................................... 52
Specifieke kenmerken ............................................................................................................................................. 52
Pathogenese ............................................................................................................................................................ 52
Pathogeen vermogen .............................................................................................................................................. 52

4

, Staal ......................................................................................................................................................................... 52
Isolatie en identificatie ............................................................................................................................................ 52
H9. Niet ........................................................................................................................................................................... 52
H10. Gram-negatieve bacillen die speciale groeivoorwaarden vereisen ........................................................................ 53
10.1. Geslachten en algemene eigenschappen .......................................................................................................... 53
10.2. Brucella spp. ....................................................................................................................................................... 53
Kenmerken .............................................................................................................................................................. 53
Pathogenese ............................................................................................................................................................ 53
Pathogeen vermogen .............................................................................................................................................. 53
Staal ......................................................................................................................................................................... 53
Isolatie en identificatie ............................................................................................................................................ 53
10.3. Heamophillus influenzae .................................................................................................................................... 54
Algemene eigenschappen en species Heamophillus spp........................................................................................ 54
Specifieke kenmerken ............................................................................................................................................. 54
Pathogenese ............................................................................................................................................................ 54
Pathogeen vermogen .............................................................................................................................................. 54
Staal ......................................................................................................................................................................... 54
Isolatie en identificatie ............................................................................................................................................ 54
10.4. Legionella ........................................................................................................................................................... 55
Specifieke kenmerken ............................................................................................................................................. 55
Pathogenese ............................................................................................................................................................ 55
Pathogeen vermogen .............................................................................................................................................. 55
Staal ......................................................................................................................................................................... 55
Isolatie en identificatie ............................................................................................................................................ 55
10.5. Bordetella pertussis ........................................................................................................................................... 56
Kenmerken .............................................................................................................................................................. 56
Pathogenese ............................................................................................................................................................ 56
Pathogeen vermogen .............................................................................................................................................. 56
Staal ......................................................................................................................................................................... 56
Isolatie en identificatie ............................................................................................................................................ 56
10.6. Campylobacter spp. ........................................................................................................................................... 56
Kenmerken .............................................................................................................................................................. 56
Pathogenese ............................................................................................................................................................ 56
Pathogeen vermogen .............................................................................................................................................. 56
Staal ......................................................................................................................................................................... 56
Isolatie en identificatie ............................................................................................................................................ 56
10.7. Helicobacter pylori ............................................................................................................................................. 57
Kenmerken .............................................................................................................................................................. 57
Pathogenese ............................................................................................................................................................ 57

5

, Pathogeen vermogen .............................................................................................................................................. 57
Staal ......................................................................................................................................................................... 57
Isolatie en identificatie ............................................................................................................................................ 57
H11. Antibiogram ............................................................................................................................................................ 58
11.1 Antibiogram......................................................................................................................................................... 58
11.2 Gevoeligheid van bacteriën................................................................................................................................. 58
11.3 Antibiogram volgens de dilutiemethode............................................................................................................. 59
Dilutie in vloeibaar midden ..................................................................................................................................... 59
Plaatdilutie methode............................................................................................................................................... 59
11.4 Antibiogram volgens de diffusiemethode ........................................................................................................... 59
Beïnvloedende factoren .......................................................................................................................................... 59
Kirby-Bauer methode .............................................................................................................................................. 60
11.5. Neo-Sensitabs methode & kwaliteitscontrole ................................................................................................... 60
11.6. Keuze van antibiotica ......................................................................................................................................... 60
11.7. Epsilometer of E-test .......................................................................................................................................... 61
11.8. Frequent voorkomende stammen ..................................................................................................................... 61
11.9. De 90/60 regel.................................................................................................................................................... 61
11.10. Intrinsieke resistentie ....................................................................................................................................... 61
H12. Antibiotica............................................................................................................................................................... 62
12.1. Begrippen ........................................................................................................................................................... 62
Antimicrobiële middelen ......................................................................................................................................... 62
Soorten antibiotica .................................................................................................................................................. 62
12.2. Algemene werkingsmechanismen ..................................................................................................................... 62
12.3. Farmacokinetiek ................................................................................................................................................. 62
12.4. Spectrum van antimicrobiële activiteit .............................................................................................................. 63
12.5 Algemene resistentiemechanismen .................................................................................................................... 63
12.6 Indeling van de antibiotica .................................................................................................................................. 64
12.6.1 Remming celwandsynthese.......................................................................................................................... 64
12.6.2 Remming van eiwitsynthese ........................................................................................................................ 67
12.6.3 Remming van het nucleïnezuurmetabolisme .............................................................................................. 68
12.6.4 Functie van het membraan verstoren .......................................................................................................... 69
12.7. Keuze van antibioticum ...................................................................................................................................... 69
12.8. Resistentie tegen antibiotica bij bacteriën ......................................................................................................... 69
H13. Methoden om microbiële groei tegen te gaan ....................................................................................................... 70




6

,H0. Inleiding
0.1. Pathogene micro-organismen
Taxonomie in de bacteriologie

1. Terrein van de medische bacteriologie
2. Pathogene µorganismen
3. Classificatie van µorganismen
a. Morfologische classificatie
b. Biochemisch classificatie
c. Numerische classificatie
d. Fylogenetische classificatie

1. Terrein van de medische bacteriologie
= µorganismen die zich aangepast hebben aan de mens

a. Microbiota: normaal onschadelijk
b. Transiënte omgevingsflora: regelmatig terug te vinden en normaal onschadelijk
c. (opportunistische) pathogenen: schadelijk → virulentiefactoren!

2. Pathogene µorganismen
→ infectieziekten

a. Virussen
b. Bacteriën prokaryote ééncelligen
c. Schimmels eukaryote meercelligen
d. Gisten eukaryoten ééncelligen
e. Parasieten
a. Protozoa eukaryote ééncelligen
b. Metazoa eukaryote meercelligen: helminthes, arthropoda

Prokaryote cel Eukaryote cel




Size 5-10µm 0.5-2.0µm
Nucleus + -
Organellen + -
Celwand - +



3. Classificatie van µorganismen
a. Morfologisch
b. Biochemisch
c. Numerische
d. Fylogenetische




7

,0.2. Morfologie en kokken
Morfologische classificatie
- Gramkleuring → vorm, kleur en
groepsverband
o Gram +: paars
o Gram -: roze/rood
- Microscopische vorm
o Bacil – staaf
o Kok – rond



Filamenteuze bacteriën
Drie geslachten

a. Actinomyces gram +
niet zuurvast
anaëroob
b. Nocardia gram +
zuurvast
aëroob
c. Streptomyces gram +
niet zuurvast
aëroob

Echte bacteriën
Kokken
- Streptokokken gram + rijen (ketens)
- Stafylokokkken gram + trossen
- Tetraden per vier, +/- in vierkant
- Sarcinae gram + 3D groepen van 8 cellen
- Neisseriae gram - boonvormig
- Veillonelle gram - (diplo)coccen


Bacillen
- Zuurvast mycobacterium tuberculosis
- Gram + sporevormers bacillus (aëroob)
clostridium (anaëroob)
- Gram + niet sporevormers corynebacterium
lactobacillus
listeria
- Gram – staven: enterobacteriaceae
pseudomonas

Vibrio’s (gram -)


Spirillen (gram -)
- Gebogen, komma vorm

Spirocheten
- Borrelia burgdorfero (gram -)
- Treponema pallidum (gram -)
- Leptospira interrogans
8

,0.3. Taxonomie
Biochemische classificatie
• Reincultuur: d.i. een cultuur van bacteriën die allemaal afstammen van 1 cel, een geïsoleerde kolonie

• Species: = soort: kleinste groepering van bacteriën met dezelfde eigenschappen (=naam):
➔ Identificatie tot species-niveau

• Stammen: culturen van een soort, primair geïsoleerd uit hun natuurlijke omgeving, die geen onderling
verband hebben (nummer: ATCC): 2 verschillende stammen met zelfde naam en
verschillende nummer
➔ Typeren van een stam (serotypering, moleculair, biochemisch,…)

• ATCC: American Type Culture Collection: verzameling stammen

• Biotypen of variëteiten: biotypen E. coli verschillen van elkaar in een kleinere, minder belangrijke
biochemische reactie (verschillende stammen). Zo ook serotypes

• Isolaat: men kan bij leden van hetzelfde gezin verschillende isolaten van één bepaalde stam van een
bacterie isoleren

• Serotypen: serotypen van Salmonella vb. verschillen van elkaar in celwandantigenen
• Stam geserotypeerd indien al biochemisch geïdentificeerd
o Salmonella drie soorten antigenen kunnen bezitten
▪ O-antigenen → polysachariden= deel v. celwand + thermostabiel
▪ H-antigenen → proteïnen = deel v. flagella + thermolabiele
▪ Vi-antigenen → in buitenste lagen en maskeert O-antigen

• Bergey’s manual: dit boek geeft een overzicht van de bacteriën ingedeeld volgens het biochemisch
classificatiesysteem

➔ Species, Genus, Familie, Orde, Klasse, Afdeling, Rijk


Numerische classificatie
- Iedere eigenschap evenveel belang
- Adanson: G = Ng x 100 / Ng + Nv
o G = gelijkende index tussen 2 stammen
o Ng = #gemeenschappelijk kenmerken
o Nv = #kenmerken waarin ze verschillen

Fylogenetische classificatie
- GC – AT verhouding bepalen
o Als bacteriën fylogenetisch dicht bij elkaar:
GC – AT verhouding ook dicht bij elkaar
- Volgens evolutieve verwantschap
- Kijken naar evolutieve verhouding tussen bacteriën
onderling




9

, 0.4. De celwand van de bacterie




Gram-positief Gram-negatief
Kleur Paars Roze
Representatieve genera Bacillus, Staphylococcus, Escherichia, Neisseria,
Streptococcus Pseudomonas
Aanwezige structuren/componenten
- tratglycaan Dikke laag Dunne laag
- Teochoic acids
- Periplasm
- Buitenste membraan
- Endotoxin (lipopolysaccharide)
- Porin proteins (onnodig want geen buiten membraan) (toestaan passage van moleculen door buitenste
membraan)
Algemene karakteristieken
- Gevoeligheid aan penicilline Algemeen meer gevoelig (met Algemeen minder gevoelig (met
uitzonderingen!) uitzonderingen!)
- Gevoeligheid aan lysozyme
- Form dat resulteert in verwijdering Protoplast Spheroplast
van peptidoglycaan




10

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