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Immunology > Pregnancy Test and ELISA Test > Flashcards

Pregnancy Test and ELISA Test Flashcards

1
Q

Checks your pee or blood for a hormone called human chorionic gonadotropin (hCG)

A

Pregnancy Test

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2
Q

The body makes this hormone after a fertilized egg attaches to the wall of the uterus

A

Human Chorionic Gonadotropin (hCG)

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3
Q

The increase of this hormone usually happens about 6 days after fertilization. It rises quickly, doubling every 2 to 3 days

A

Human Chorionic Gonadotropin

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4
Q

Types of Pregnancy Test

A

Blood Tests and Urine Tests

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5
Q

You can take these at home or in a doctor’s office. Along with being private and convenient, this type of test is quick and easy to use. It is also very accurate if the directions are followed properly.

A

Urine Tests/Home Pregnancy Tests

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6
Q

Ways to Perform Urine Tests/Home Pregnancy Tests

A
  • Hold the test stick in your urine stream
  • Collect pee in a cup and dip the test stick into it
  • Collect pee in a cup and use a dropper to put it into another container
  • Wait for a few minutes before seeing the results
  • After taking the test, confirm the results with a doctor who can perform more sensitive tests
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7
Q

This type of pregnancy test is performed at the doctor’s office. It can detect pregnancy earlier than a home pregnancy test, about 6 to 8 days after ovulation.

Takes longer to get the results than with a home pregnancy test

A

Blood Tests

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8
Q

Two Types of Blood Pregnancy Tests

A

Qualitative hCG Test and Quantitative hCG Test

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9
Q

Simply checks for hCG- It gives a “yes” or “no” answer to the question, “Are you
pregnant?“
• Doctors often order these tests to confirm pregnancy as early as 10 days after conception.
• Some can detect hCG much earlier.

A

Qualitative hCG Test

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10
Q

measures the exact amount of hCG in your blood.
• It can find even very low levels of hCG.
• These tests may help track problems during pregnancy.
• Your doctor may use them along with other tests to rule out an ectopic pregnancy, when the fertilized
egg implants outside your uterus, or after a miscarriage, when hCG levels fall quickly.

A

Quantitative hCG Test (beta hCG)

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11
Q

A quantitative immunological procedure in which Ag-Ab reaction is monitored by enzyme measurements. It is highly sensitive

A

Enzyme-linked Immunosorbent Assay (ELISA)

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12
Q

Samples routinely used in ELISAs

A

Serum, plasma, cell culture supernates, cell lysates, saliva, tissue lysates, and urine

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13
Q

An ELISA test may be used to diagnose:

A

HIV, Lyme Disease, Pernicious Anemia, Rotavirus, Syphilis, and others

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14
Q

First to label antibodies with a fluorescent dye, and use it to identify antigens in tissue sections. This method is known today as immunofluorescence.

A

Albert H. Coons and his colleagues (1941)

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15
Q

Described radioimmunoassay

A

Rosalyn Sussman and Soloman Berson (1960)

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16
Q

Independently invented the ELISA test

A

Eva Engvall and Peter Perlman (1971)

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16
Q

A method in which a conjugated substrate competes with a protein of interest. Developed and used to detect hCG

A

Competitive ELISA Method (1976)

17
Q

A method in which the detection antibody is coated onto the plate surface before the protein of interest is added; is developed and tested on several substrates for proof-of-concept

A

Sandwich ELISA Method (1977)

18
Q

A method in which a secondary antibody is added for detection purposes; developed and used to detect human serum albumin

A

Indirect ELISA (1978)

19
Q

What screening test was first used for testing HIV? Give the exact date as well

A

ELISA Test - March 2, 1985

20
Q

Discuss the principle behind ELISA

A
  • Uses an enzyme to detect the binding of Ag-Ab
  • The enzyme converts a colorless substrate to colored product
  • That indicates the presence of Ag
  • It can be used to detect the presence of both Ag-Ab
21
Q

The Four Main Types of ELISAs

A
  • Direct
  • Indirect
  • Sandwich
  • Competitive
22
Q

When to use Direct ELISA?

A

Assessing antibody affinity and specificity.

Investigating blocking/inhibitory interactions.

23
Q

Advantage of Direct ELISA

A

It is a fast and simple protocol

24
Q

Disadvantages of Direct ELISA

A
  • Less specific since you are only using 1 antibody.

* Potential for high background if all proteins from a sample are immobilized in well.

25
Q

When to use Indirect ELISA

A

Measuring endogenous antibodies

26
Q

Advantages of Indirect ELISA

A
  • Amplification using a secondary antibody
  • Cost Effective
  • Different visualization markers can be used with the same primary antibody
27
Q

Disadvantages of Indirect ELISA

A
  • Potential for cross reactivity caused by secondary antibody
  • An extra incubation step is required in the procedure.
28
Q

When to use Sandwich ELISA

A

Determining analyte concentration in a biological sample.

28
Q

When to use Sandwich ELISA

A

Determining analyte concentration in a biological sample.

29
Q

When to use Sandwich ELISA

A

Determining analyte concentration in a biological sample.

30
Q

Advantages of Sandwich ELISA

A
  • Highest specificity and sensitivity
  • Compatible with complex sample matrices
  • Flexibility and sensitivity, since both direct and indirect detection methods can be used.
31
Q

Disadvantages of Sandwich ELISA

A
  • Longer protocol

* Challenging to develop

32
Q

When to use Competitive ELISA

A

Determining concentrations of a small molecules and hormones.

33
Q

Advantages of Competitive ELISA

A
  • Ability to quantitate small molecules
  • High specificity, since two antibodies are used.
  • High sensitivity, since both direct and indirect detection methods can be used.
  • Suitable for complex samples, since the antigen does not require purification prior to measurement.
34
Q

Disadvantages of Competitive ELISA

A
  • Less specific since you are only using 1 antibody

* Requires a conjugated antigen

35
Q

The first 4 steps in ELISA

A

Step 1
Capture antibody binds to ELISA plate wells
Step 2:
Add sample to well antigen within the sample binds to the capture antibody.
Step 3:
Wash microplate Unbound material is washed away, leaving only the antigen of interest
Step 4:
Add detection antibody Enzyme conjugated detection antibody binds to a second site on the antigen of interest

36
Q

The second 4 steps in ELISA

A

Step 5:
Wash microplate Unbound antibodies are washed away, leaving only those specific for the target of interest
Step 6:
Add substrate Substrate is converted by the enzyme on the detection antibody, producing a color change
Step 7
Read plate The microplate reader detects the colored reaction product and outputs optical density (OD) values
Step 8:
Calculate results The amount of antigen in each sample is calculated and analyzed

37
Q

Determines the presence of antigen and/or antibody; Positive or Negative Result

A

Qualitative Test

38
Q

Determines the quantity of antibody; No. of positive and negative result

A

Quantitative Test

39
Q

• It is a good techniques and being popular today because of its simple and not
involve radiation .
• It is easy than other test .
• It is most useful than other test because of their sensitivity.
• It is not harmful.

A

ELISA

Immunology (3 decks)

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