Eli

Question 1

Order of Leukocyte Frequency

Answer 1

• Neutrophils
• Lymphocytes
• Monocytes
• Eosinophils
• Basophils

Never Let Monkeys Eat Burritos

Question 2

Neutrophil

Appearance & Function

Answer 2

Granulocyte
Multiple nuclei & light granules
Phagocytosis

Question 3

Eosinophil

Appearance & Function

Answer 3

Granulocyte
Red/ orange granules
Helminth defense

Question 4

Basophil

Appearance & Function

Answer 4

Granulocyte
Dark granules
Allergic reactions

Question 5

Monocyte

Appearance & Function

Answer 5

Agranulocyte
One bean shaped nucleus
Phagocytosis

Question 6

Lymphocyte

Appearance & Function

Answer 6

Agranulocytes
Small and dark (same size as RBCs)
Specific immunity

Question 7

Serology

Answer 7

use of antigen-antibody interactions to diagnose disease

Question 8

Antigen

Answer 8

component of pathogen that stimulates the

immune system

Question 9

Antibody

Answer 9

proteins made by B-cells that bind

to the antigen

Question 10

ELISA

Answer 10

Enzyme linked immnosorbent assay

Question 11

Direct ELISA

Answer 11

Antigen (pathogen): Patient sample

Antibody (patient serum): Produced in the lab

Question 12

Indirect ELISA

Answer 12

Antigen (pathogen): Produced in the lab

Antibody (patient serum): Patient sample

Question 13

ELISA Steps

Answer 13

1. Antigen is coated onto the test plate
2. Primary antibody (from patient serum) is added
3. Secondary antibody (anti-human; has an enzyme tagged on it) is added
4. Substrate for enzyme is added
5. (+) = color change

Question 14

If a patient doesn’t have the primary antibody, will a color change occur?

Answer 14

No

Question 15

If a positive color change occurs, does that confirm that the patient has disease?

Answer 15

No. A positive ELISA only indicates that the patient has antibodies against a particular pathogen.

Question 16

Lysozyme

Answer 16

enzyme that destroys bacterial peptidoglycan by
targeting the NAG-NAM connections
• Part of the innate immune system
• Found in tears, saliva, and sweat

Question 17

What media do you use for Lysozyme and Tears?

What was the process like?

Answer 17

NA Plates

in 6 different dilutions like pizza slices

Question 18

What media do you use for saliva?
What was the process like?
Pathogen?

Answer 18

Blood agar plate
4 quadrant swabing a lawn on one
then streaking w/loop on the others
creating isolation

Pathogen: Streptococcus pygoenes

Question 19

What media do you use for microbes of the skin?

What was the process like?

Answer 19

MSA Plate
4 quadrant swabing a lawn on one
then streaking w/loop on the others
creating isolation

Pathogen: Staphylococcus aureus microbes of the skin

Question 20

Does lysozyme affect G+ bacteria, G- bacteria, or both?

Answer 20

G+ bacteria

Question 21

Why do you think lysozymes are more effective against

G+ bacteria?

Answer 21

Lysozymes destroy peptidoglycan cell wall

Question 22

Blood agar plate

Answer 22

Selective: nothing
Differential: hemolysis

Gamma = no hemolysis 
Alpha = partial hemolysis 
Beta = complete hemolysis

Pathogen: Streptococcus pyogenes = Beta hemolytic

Question 23

Mannitol salt agar (MSA) plate

Answer 23

Selective: halophiles (growth)
Differential: mannitol fermentation (positive = yellow)

Pathogen: Staphylococcus aureus = positive for mannitol
fermentation

Question 24

Limitations of the Elisa test for patient diagnosis?

Answer 24

1. a positive result only confirms there is an antibody present. but it does not mean the patient is currently ill.
2. People who are poor producers of antibodiesor who have an interfering substance may get a false positive because the antibody amount is too low and can go undetected.

Question 25

What are the main reagents added to the ELISA test.

Answer 25

1. ANTIGEN 2. PRIMARY ANTIBODY 3. SECONDARY ANTIBODY 4. SUBSTRATE

Question 26

ANTIGEN

Answer 26

Component of a *pathogen* that will stimulate the immune system. Coats the wells.

Question 27

PRIMARY ANTIBODY

Answer 27

Patients serum antibodies. If antibodies against the antigen are present then they will bind to the antigen that is at the bottom of the wells.

Question 28

SECONDARY ANTIBODY

Answer 28

Animal antibody solution. Antibody against human antibodies which is covalently conjugated to an enzyme.

Question 29

SUBSTRATE

Answer 29

Colorgenic enzyme substrate solution. Interacts with the enzyme on the second antibody which generates a visible color.

Question 30

What is the purpose of having a negative control in ELISA?

Answer 30

Incase enzymes they stick to the plastic sides and create a false positive.

Question 31

Why is is necessary to rinse sample wells between reagent steps?

Answer 31

Rinsing will remove the antibodies that did not react with the antigen. And you can get a false positive if not rinsed.

Question 32

What should you observe as a positive result for the ELISA test? What does this tell you about the patient?

Answer 32

A positive result will result in a color which tells us the patient has antibodies

Question 33

Initial HIV testing is performed via an ELISA | Which immunologic method is used as the verification test for a positive ELISA HIV result?

Answer 33

Western blot test | Separates proteins by electrophoresis and uses antibodies to detect proteins.