Systems of Pathogen Detection I Flashcards
Why is taxonomy better than standard naming for pathogens?
Names only imply capacity for pathogenicity
They do not include an assessment of virulence
Outline the taxonomy of TB
Domain Eubacteria Kingdom Procaryote Phylum Gram positive Class Actinobacteria Order Actinomycetales Family Mycobacteriaceae Genus Mycobacterium Species Mycobacterium tuberculosis Strain Beijing Type IIa6B1 Isolate Sputum
Of the myobacterium genus which are pathogenic to humans?
148 current species - Only three are obligate human pathogens:
- M .tuberculosis
- M .leprae
- M .ulcerans
Define a pathogen
A microbe capable of causing a specific degree of host damage
What is a commensal non pathogen (in host)?
PRESENT but NOT CAPABLE of causing disease in the host
eg. E.coli Bacteroides thetaiotaomicron
‘good bacteria’
What is a Zoonotic Non pathogen (in carrier)?
PRESENT but only CAPABLE of causing disease in ANOTHER host
eg. E.coli O157:H7 is subclinical in cattle
What is a commensal opportunist (in host)?
PRESENT and CAPABLE of causing disease in the host but only in certain circumstances
eg. Bacteroides fragilis Coagulase Negative Staphylococcus (CNS)
Why does a positive sample not confirm disease?
Being infected / presented with disease doesn’t guarantee active disease
Majority people presented with a pathogen don’t develop infection at all or it becomes latent and activated later on in life
What determines how good a test result is?
A test result is only as good as the sample provided
Describe the requirements of a sterile testing site
Sterile sites must be free from contamination
e.g. Skin flora in blood cultures
How must non-sterile sites of testing be prepared?
Non sterile sites require decontamination of normal flora
e.g. Faeces, Mouth, Skin
Why are some samples concentrated?
Samples with high volume or relatively low infected pathogen load require concentration (centrifugation, filtering)
e.g. CSF, Ascites, 24 hr Urine
Why do we culture pathogens?
Culturing the bacteria proves it’s presence
But don’t always require culturing but knowing which specific pathogen you’re looking for makes identification a lot easier
How is a culture prepared?
enrichment
purification
amplification
How is a direct sample prepared for identification?
concentrated and treated
What identification techniques are used to identify pathogens in a sample?
Molecular DNA/RNA
Microscopy: gross morphology
HPLC MassSpec: chemical Composition
Which pathogens are visible via direct light microscopy?
Larger pathogens
- Trichomonas vaginalis
- Schistosoma mansoni
- Entamoeba histolytica
- Strongyloides (thread worm)
When is Electron microscopy used?
Smaller sized samples
Mainly used for viruses
Used when PCR and culturing isn’t an option
> not used for diagnostics
Which pathogens are viewed using electron microscopy?
- Rotavirus (faeces)
- Rabies lyssavirus (brain tissue)
- Hepatitis B (liver)
- Tonsilitis adenovirus - nasal secretion
When are samples stained?
Stain direct bacterial samples to identify using light microscopy
What does staining show us?
Can identify:
- Gram stain
- Shape
- Features (flagella etc.)
- Gram +/-
What is the significance of bacterial staining?
We can also use this staining to identify capsulated organisms as these are more likely to be pathogens
Give examples of capsulated bacteria
- streptococcus pneumoniae
Bacterial spores also present in Bacillus anthracis and Clostridium perfringens (food poisoning)
How is immunofluorescent staining done?
Immunofluorescent staining with pathogen specific conjugated antibody