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Elisa > Eli > Flashcards

Eli Flashcards

1
Q

Order of Leukocyte Frequency

A
  • Neutrophils
  • Lymphocytes
  • Monocytes
  • Eosinophils
  • Basophils

Never Let Monkeys Eat Burritos

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2
Q

Neutrophil

Appearance & Function

A

Granulocyte
Multiple nuclei & light granules
Phagocytosis

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3
Q

Eosinophil

Appearance & Function

A

Granulocyte
Red/ orange granules
Helminth defense

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4
Q

Basophil

Appearance & Function

A

Granulocyte
Dark granules
Allergic reactions

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5
Q

Monocyte

Appearance & Function

A

Agranulocyte
One bean shaped nucleus
Phagocytosis

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6
Q

Lymphocyte

Appearance & Function

A

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

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7
Q

Serology

A

use of antigen-antibody interactions to diagnose disease

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8
Q

Antigen

A

component of pathogen that stimulates the

immune system

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9
Q

Antibody

A

proteins made by B-cells that bind

to the antigen

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10
Q

ELISA

A

Enzyme linked immnosorbent assay

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11
Q

Direct ELISA

A

Antigen (pathogen): Patient sample

Antibody (patient serum): Produced in the lab

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12
Q

Indirect ELISA

A

Antigen (pathogen): Produced in the lab

Antibody (patient serum): Patient sample

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13
Q

ELISA Steps

A
  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
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14
Q

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

A

No

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15
Q

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

A

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

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16
Q

Lysozyme

A

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

17
Q

What media do you use for Lysozyme and Tears?

What was the process like?

A

NA Plates

in 6 different dilutions like pizza slices

18
Q

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

A

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

Pathogen: Streptococcus pygoenes

19
Q

What media do you use for microbes of the skin?

What was the process like?

A

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

20
Q

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

A

G+ bacteria

21
Q

Why do you think lysozymes are more effective against

G+ bacteria?

A

Lysozymes destroy peptidoglycan cell wall

22
Q

Blood agar plate

A

Selective: nothing
Differential: hemolysis

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

Pathogen: Streptococcus pyogenes = Beta hemolytic

23
Q

Mannitol salt agar (MSA) plate

A

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

Pathogen: Staphylococcus aureus = positive for mannitol
fermentation

24
Q

Limitations of the Elisa test for patient diagnosis?

A
  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.
25
Q

What are the main reagents added to the ELISA test.

A
  1. ANTIGEN
  2. PRIMARY ANTIBODY
  3. SECONDARY ANTIBODY
  4. SUBSTRATE
26
Q

ANTIGEN

A

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

27
Q

PRIMARY ANTIBODY

A

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.

28
Q

SECONDARY ANTIBODY

A

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

29
Q

SUBSTRATE

A

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

30
Q

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

A

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

31
Q

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

A

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

32
Q

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

A

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

33
Q

Initial HIV testing is performed via an ELISA

Which immunologic method is used as the verification test for a positive ELISA HIV result?

A

Western blot test

Separates proteins by electrophoresis and uses antibodies to detect proteins.

Elisa (1 decks)

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