Immunoassays and Automated Procedures

Question 1

Which Immunoassay step requires washing steps?

Answer 1

Heterogeneous Immunoassays
Solid Phase
Competitive/Noncompetitive

Question 2

Which Immunoassay step does NOT require a washing phase?

Answer 2

Homogeneous Immunoassays
Liquid Phase
Competitive
Faster/Easier

Question 3

What are the different labels that are applied in immunoassays?

Answer 3

Enzyme Immunoassay
Chemiluminescence
Fluorescent Substances

Question 4

What is Chemiluminescence?

Answer 4

Light emission produced during a chemical reaction

Question 5

What are ELISA or EIA designed to do?

Answer 5

Detect antigens/antibodies by producing an enzyme triggered color change

Question 6

What are the general approaches of EIA?

Answer 6

Nonisotopic labels (Detect total IgM/IgG)
Catalytic properties of enzymes (Detects and quantitates reactions)

Question 7

What qualifications must be met for an enzyme to be used in immunoassay? (NOT SAUCE)

Answer 7

High stability
Extreme specificity
Not be present in antigen/antibody during testing

Question 8

How do we quantify a Noncompetitive Enzyme Immunoassay?

Answer 8

Amount of color change is proportional to amount of antibody

Question 9

How do we quantify a Competitive Enzyme Immunoassay?

Answer 9

Amount of color change is inversely proportional to amount of antibody

Question 10

What are Capture Enzyme Immunoassays designed to detect?

Answer 10

Specific antibody type
IgM
Toxoplasma IgM

Question 11

What are Chemiluminescent labels used to detect?

Answer 11

Proteins
Viruses
Oligonucleotides
Genomic nucleic acid sequences
Competitive or Sandwich

Question 12

How do we quantify a Capture Enzyme Immunoassay?

Answer 12

Amount of light emitted is inversely proportional to amount of analyte (antigen)

Question 13

What can luminol, acridinium esters, peroxyoxaltes, dioxetanes, and tris rutenium be grouped into?

Answer 13

Chemiluminescent labels

Question 14

What has been used as visual tags for the presence of antibody?

Answer 14

Conjugated enzyme labels:
HRP
ALP
Avidin-Biotin

Question 15

Which assay uses a conjugated antibody to detect antigen-antibody reactions at a microscope level?

Answer 15

Direct Immunofluorescent Assay (DFA)

Question 16

Which assay is used to confirm the specificity of the FA technique?

Answer 16

Inhibition Immunofluorescent assay (IIA)

Question 17

What is used in detecting Listeria Monocytogenes?

Answer 17

SQUID Technology

Question 18

What test is measured photometrically and is assessed by the amount of cloudiness or turbidity?

Answer 18

Nephelometry

Question 19

What is Nephelometry used to measure?

Answer 19

Complement components
Immune complexes
Presence of antibodies

Question 20

What is Nephelometry based on?

Answer 20

The reaction between the protein being assayed and a specific antiserum

Question 21

What is known as the relationship between antigen and the measurement of a constant antibody concentration?

Answer 21

Heidelberger Curve

Question 22

What is Flow Cell Cytometry based on?

Answer 22

Reaction of dyes with the cellular component of interest
Stained cells pass through laser

Question 23

What is the Polymerase Chain Reaction (PCR)?

Answer 23

In vitro method that amplifies low levels of DNA

Question 24

What are the three cycles in PCR?

Answer 24

DNA Denaturation (Separation)
Primer Annealing (Recombination)
Extension of DNA (Synthetization)

Question 25

What is a disadvantage of PCR?

Answer 25

Prone to producing false positives Highly Sensitive (good thing)

Question 26

What are three important applications of PCR?

Answer 26

DNA amplification Target sequence Identification Labeled antisense probe synthesis

Question 27

What can PCR analysis lead to?

Answer 27

Detection of gene mutations Coronary artery disease

Question 28

What kind of charge does DNA have?

Answer 28

Net Negative charge Migrates towards the anode (positive)

Question 29

What are the advantages of molecular genetic testing?

Answer 29

Faster turnaround time Smaller sample size Increased specificity and sensitivity

Question 30

What is considered the gold standard of molecular methods?

Answer 30

DNA sequencing Displays exact nucleotide sequence

Question 31

Why is the Sanger method important?

Answer 31

It is the most frequently used DNA sequencing method

Question 32

What is a modified PCR technique that uses fluorescence resonance energy transfer to quantitate DNA sequences?

Answer 32

Real-Time PCR

Question 33

What is a modified PCR technique that is used in detection and quantification of HIV and CMV?

Answer 33

Nucleic Acid Sequence Amplification

Question 34

Which blot technique uses specimen DNA that are separated by electrophoresis?

Answer 34

Southern Blot Sickle Cell Anemia is seen

Question 35

Which blot technique uses mRNA from the specimen and is separated by electrophoresis?

Answer 35

Northern Blot

Question 36

Which blot technique separates proteins electropheretically?

Answer 36

Western Blot