Identification and Diagnosis Flashcards
What are the five “I’s” of culturing microrganisms?
- Inoculate
- Incubate
- Isolate
- Inspect
- Identify
What does it mean to:
- Inoculate
- Incubate
- Place into a nutrient rich media
- Place inoculum into ideal enviroment
What are the three types of idenitification methods for grown cultures?
- Phenotypic methods
- Direct and indirect methods
- Immunological methods
- Genotypic (genetic) methods
What are phenotypic analyses direct methods?
- Staining
- Cultivation of specimen
What are these types of stains:
- Dye stains all microrganisms the same color
- Dye attaches to microgorganism, stains certain color
- Simple Stain
- Positive stain
Name these stains:
- Dye stains everything around microorganism
- Use of multiple stains to distinguish cell types
Negative stain
Differential stain
What are the two common types of differential stains?
- What do they do?
-
Gram Stain
- Checks for Gram +/-
-
Acid Fast Stain
- Checks for lipid/non-lipid walls
Name this stain type:
Stain that identify the specific structure of bacteria
Structural Stain
What are the types of Structural Stains?
- What do they do?
-
endospore stain
- Spore vs. vegetative cell
-
Flagellar stain
- Checks for flagella, number and location
-
Capsular stain
- checks for capsule
Name these types of cultivation methods:
- Used to grow variety of bacteria, no special growth factors
- Has additional nutrients for bacteria to grow better.
- General media
- Enriched media
Bacteria that require enriched media to grow are called:
Fastidious
Name these cultivation methods:
- Allow certain bacteria to grow, inhibits others
- Used to identify two closely related bacteria
- Selective media
- Differential media
Name these cultivation methods:
- used to identify anaerobic bacteria/oxygen requirments
- Provides stable conditions for later analysis
- Reducing media
- Transport media
What are the different types of media?
- Liquid
- Semisolid
- Solid
What are examples of:
- General media
- Enriched media
- Nutrient agar
- Chocolate agar
Give examples of:
- Selective media
- Differential media
What do they grow?
- EMB agar media
- gram negative only
- Blood agar media
- check for alpha, beta, and gamma bacteria
What is an example of both a selective and differential media?
What does it grow?
Mannitol salt agar
- selective for staphococcus
- differential for staph species
What are the two ways to get a pure culture?
- Streak plate to isolate individual bacteria so they grow to form a colony
- Use differential/selective media
What is phage typing?
indirect examination method
Bacteriaphages are distributed to different sections of agar plates.
Phages only lyse certain bacteria.
What is antimicrobial sensitivity (examination method)?
- What effects does it have?
An indirect method
Different antimicrobials are placed on agar plate.
- Clear zones= susceptible
no zone= resistant
What is the disc diffusion method of antimicrobial sensitivity?
Discs impregnated with antibiotics are placed on agar.
What is biochemical testing (examination method)?
An indirect method
Evidence that finds particular factor present with particular bacteria.
How does Biochemical testing work?
- What are the different reactions that can occur?
Bacteria are inoculated in substrate containing media and test for end product.
- reaction = enzyme present
- reaction= no enzyme
What are these different biochemical tests:
- Produces enzyme to ferment sugars
- Produces enzyme to break down gelatin
- Produces enzyme to break down starch
- Carbohydrate fermentation
- Gelatin hydrolysis
- Starch hydrolysis
What are:
- Antigens
- Antibodies
- Anything that stimuates an immune response by B or T cells
- immunce cels from your body (specific for each antigen)
What is immunological analysis?
- How can this test vary in specicifity and sensitivity?
Blood test that analyzes antigen and antibody reactions.
- Specifically= Reacts w/ specific antibody/antigen
- Sensitivity= Can detect very small amounts of antibody/antigen
What are the two methods for immunological assays?
- Take patient serum, add known antigen, look for reaction with antibody
- Take unknown microbe, add known antibody, look for reaction with antigen
Name these immunological analysis types:
- can determine (antigen) when antibodies attach and form clumps
- Antigen is made insoluble by an antibody, creates clody zone
- Agglutination test
- Precipitation test
Name these immunological analysis types:
- Separate bacterial proteins using electric current on a gel and expose to specific antigen
- Antibodies labeled with fluorescent dye bind antigen
- Western blot
- Immunoflourescence
Name these immunological analysis types:
- Radioactively labeled antibody/antigen
- Calorimetric test to detect antibody/antigen
- Radioimmunoassay
- ELISA
Name these immunological analysis types:
Introduce antigen/antibody into patient to produce reaction
In-vivo
What is genotypic analysis?
A PCR (polymerase chain reaction)
Use of small primer sequence to amplify a genome (compare to known genomes)
What makes genotypic analysis so effective?
Very sensitive and fast (hours)
Can analyze DNA or RNA genomes
What are these types of Genotypic analysis:
- Analysis of ribosomal RNA
- DNA probes with flourescent molucule
- Ribotyping
- DNA hybridization (FISH)
