Identification and Diagnosis

Question 1

What are the five “I’s” of culturing microrganisms?

Answer 1

1. Inoculate
2. Incubate
3. Isolate
4. Inspect
5. Identify

Question 2

What does it mean to:

1. Inoculate
2. Incubate

Answer 2

1. Place into a nutrient rich media
2. Place inoculum into ideal enviroment

Question 3

What are the three types of idenitification methods for grown cultures?

Answer 3

1. Phenotypic methods
   
   1. Direct and indirect methods
2. Immunological methods
3. Genotypic (genetic) methods

Question 4

What are phenotypic analyses direct methods?

Answer 4

1. Staining
2. Cultivation of specimen

Question 5

What are these types of stains:

1. Dye stains all microrganisms the same color
2. Dye attaches to microgorganism, stains certain color

Answer 5

1. Simple Stain
2. Positive stain

Question 6

Name these stains:

1. Dye stains everything around microorganism
2. Use of multiple stains to distinguish cell types

Answer 6

Negative stain

Differential stain

Question 7

What are the two common types of differential stains?

• What do they do?

Answer 7

1. Gram Stain
   
   1. Checks for Gram +/-
2. Acid Fast Stain
   
   1. ​Checks for lipid/non-lipid walls

Question 8

Name this stain type:

Stain that identify the specific structure of bacteria

Answer 8

Structural Stain

Question 9

What are the types of Structural Stains?

• What do they do?

Answer 9

1. endospore stain
   
   1. Spore vs. vegetative cell
2. Flagellar stain
   
   1. Checks for flagella, number and location
3. Capsular stain
   
   1. checks for capsule

Question 10

Name these types of cultivation methods:

• Used to grow variety of bacteria, no special growth factors
• Has additional nutrients for bacteria to grow better.

Answer 10

• General media
• Enriched media

Question 11

Bacteria that require enriched media to grow are called:

Answer 11

Fastidious

Question 12

Name these cultivation methods:

• Allow certain bacteria to grow, inhibits others
• Used to identify two closely related bacteria

Answer 12

• Selective media
• Differential media

Question 13

Name these cultivation methods:

• used to identify anaerobic bacteria/oxygen requirments
• Provides stable conditions for later analysis

Answer 13

• Reducing media
• Transport media

Question 14

What are the different types of media?

Answer 14

1. Liquid
2. Semisolid
3. Solid

Question 15

What are examples of:

1. General media
2. Enriched media

Answer 15

1. Nutrient agar
2. Chocolate agar

Question 16

Give examples of:

1. Selective media
2. Differential media

What do they grow?

Answer 16

1. EMB agar media
   
   1. gram negative only
2. Blood agar media
   
   1. check for alpha, beta, and gamma bacteria

Question 17

What is an example of both a selective and differential media?

What does it grow?

Answer 17

Mannitol salt agar

• selective for staphococcus
• differential for staph species

Question 18

What are the two ways to get a pure culture?

Answer 18

1. Streak plate to isolate individual bacteria so they grow to form a colony
2. Use differential/selective media

Question 19

What is phage typing?

Answer 19

indirect examination method

Bacteriaphages are distributed to different sections of agar plates.

Phages only lyse certain bacteria.

Question 20

What is antimicrobial sensitivity (examination method)?

• What effects does it have?

Answer 20

An indirect method

Different antimicrobials are placed on agar plate.

• Clear zones= susceptible

no zone= resistant

Question 21

What is the disc diffusion method of antimicrobial sensitivity?

Answer 21

Discs impregnated with antibiotics are placed on agar.

Question 22

What is biochemical testing (examination method)?

Answer 22

An indirect method

Evidence that finds particular factor present with particular bacteria.

Question 23

How does Biochemical testing work?

• What are the different reactions that can occur?

Answer 23

Bacteria are inoculated in substrate containing media and test for end product.

• • reaction = enzyme present
• reaction= no enzyme

Question 24

What are these different biochemical tests:

1. Produces enzyme to ferment sugars
2. Produces enzyme to break down gelatin
3. Produces enzyme to break down starch

Answer 24

1. Carbohydrate fermentation
2. Gelatin hydrolysis
3. Starch hydrolysis

Question 25

What are: 1. Antigens 2. Antibodies

Answer 25

1. Anything that stimuates an immune response by **​B or T cells** 2. immunce cels from your body (specific for each antigen)

Question 26

What is **immunological analysis?** * How can this test vary in **specicifity and sensitivity?**

Answer 26

Blood test that analyzes antigen and antibody reactions. * Specifically= Reacts w/ **specific** antibody/antigen * Sensitivity= Can detect **very small** amounts of antibody/antigen

Question 27

What are the two methods for immunological assays?

Answer 27

1. Take patient serum, add known antigen, look for reaction with antibody 2. Take unknown microbe, add known antibody, look for reaction with antigen

Question 28

Name these immunological analysis types: * can determine (antigen) when antibodies attach and **form clumps** * Antigen is **made insoluble** by an antibody, creates clody zone

Answer 28

* Agglutination test * Precipitation test

Question 29

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

Answer 29

* Western blot * Immunoflourescence

Question 30

Name these immunological analysis types: * **Radioactively** labeled antibody/antigen * **Calorimetric** test to detect antibody/antigen

Answer 30

* Radioimmunoassay * ELISA

Question 31

Name these immunological analysis types: Introduce antigen/antibody **into patient** to produce reaction

Answer 31

In-vivo

Question 32

What is **genotypic analysis?**

Answer 32

A PCR (polymerase chain reaction) Use of small **primer sequence to amplify a genome** (compare to known genomes)

Question 33

What makes genotypic analysis so effective?

Answer 33

Very sensitive and fast (hours) Can analyze DNA or RNA genomes

Question 34

What are these types of Genotypic analysis: * Analysis of ribosomal RNA * DNA probes with flourescent molucule

Answer 34

* Ribotyping * DNA hybridization (**FISH**)