Automation & Immunochemical Techniques

Question 1

3 phases of analytic process

Answer 1

Preanalytic, analytic, post-analytic

Question 2

The first continuous flow, single-channel, sequential batch analyzer capable of providing a single test result on approximately 40 samples/hr

Answer 2

AutoAnalyzer (AA)

Question 3

It was the first noncontinuous flow,
discrete analyzer as well as the first instrument to have random access
capabilities, whereby stat specimens could be analyzed out of sequence on an as-
needed basis.

Answer 3

Automatic Clinical

Analyzer (ACA) (DuPont [now Siemens]).

Question 4

This instrument was the first to use microsample
volumes and reagents on slides for dry chemistry analysis and to incorporate
computer technology extensively into its design and use.

Answer 4

Kodak Ektachem (now VITROS) Analyzer (now Ortho-
Clinical Diagnostics)

Question 5

The most recent milestone in chemistry analyzer development has been the
combination of chemistry and immunoassay

Answer 5

Modular analyzer

Question 6

liquids (reagents, diluents, and samples) are pumped

through a system of continuous tubing.

Answer 6

Continuous flow analyzer

Question 7

3 basic approaches with instruments

Answer 7

continuous flow,

centrifugal analysis, and discrete analysis.

Question 8

uses the force generated by centrifugation to transfer
and then contain liquids in separate cuvettes for measurement at the perimeter of
a spinning rotor.

Answer 8

centrifugal analysis

Question 9

Major advantage of centrifugal analysis

Answer 9

Batch analysis

Question 10

the separation of each sample and accompanying

reagents in a separate container.

Answer 10

discrete analysis.

Question 11

They are the most popular and versatile analyzers

Answer 11

discrete analysis.

Question 12

Light source used in Reflectance spectrometry

Answer 12

Tungsten-halogen lamp

Question 13

It has been and remains a manual process in most laboratories

Answer 13

Preparation of the sample for analysis

Question 14

The most sophisticated approach that is commonly used

today in specimen identification

Answer 14

Employing a bar code

Question 15

These slides have microscopically thin layers of dry
reagents mounted on a plastic support. The slides are approximately the size and
thickness of a postage stamp.

Answer 15

dry

chemistry slide

Question 16

Most automated wet chemistry analyzers use ________ that dip into
the reaction container for a few seconds to stir sample and reagents, after which
they return to a wash reservoir

Answer 16

stirring paddles

Question 17

causes major interference in many analyses.

Answer 17

Proteins

Question 18

In the older continuous flow systems, a _____ was the separation or
filtering module. It performed the equivalent of the manual procedures of
precipitation, centrifugation, and filtration, using a fine-pore cellophane
membrane.

Answer 18

dialyzer

Question 19

are attached from multiple remote stations where the reaction
mixtures reside to a centralized monochromator/detector unit that, in conjunction
with the computer, sequences and analyzes a large volume of light signals from
multiple reactions.

Answer 19

fiberoptic cables, or “light pipes”

Question 20

refers to automated devices and robots
integrated with existing analyzers to perform all phases of laboratory testing.
Most attention to date has been devoted to development of the front-end systems that can identify and label specimens, centrifuge the specimen and prepare
aliquots, and sort and deliver samples to the analyzer or to storage.

Answer 20

Total Laboratory Automation

Question 21

Developed RadioImmunoAssay (RIA)

Answer 21

Dr. Rosalyn Yallow

Question 22

Immunoprecipitation techniques: Gel-Passive

Answer 22

Double Diffusion (Ouchterlony technique)
Single Diffusion (Radial Immunodiffusion)

Question 23

Immunoprecipitation techniques: Gel-Electrophoresis

Answer 23

Counterimmunoelectrophoresis
Immunoelectrophoresis
Immunofixation Electrophoresis
Rocket electrophoresis

Question 24

Immunoprecipitation techniques: Soluble Phase

Answer 24

Turbidimetry & Nephelometry

Question 25

The Ag diffuses from the well in all directions, binds to the soluble Ab in the agarose, and forms a complex seen as a concentric precipitin ring. The diameter of the ring is related to the concentration of the Ag that diffused from the well.

Answer 25

Radial Immunodiffusion (RID)

Question 26

2 variations in RID

Answer 26

Fahey-McKelvey (kinetic) & Mancini (endpoint)

Question 27

an immune precipitation method that | uses an electrical field to cause the Ag and Ab to migrate toward each other.

Answer 27

Counterimmunoelectrophoresis

Question 28

2 methods used in the clinical lab to characterize monoclonal proteins in serum & urine

Answer 28

Immunoelectrophoresis (IEP) and immunofixation electrophoresis (IFE)

Question 29

In this quantitative technique, reagent Ab is mixed with agarose; Ag is placed in the well and electrophoresed. As the Ag moves through the agarose, it reacts with the reagent Ab and forms a “rocket,” with stronger precipitation along the edges.

Answer 29

rocket technique (Laurell technique, or electroimmunoassay)

Question 30

enhances the Ag–Ab interaction in both turbidimetry & nephelometry

Answer 30

Polyethylene glycol (PEG)

Question 31

If the binding | reagent is an Ab, it is an ______

Answer 31

immunoassay

Question 32

If the binding | reagent is a receptor, it is a _____

Answer 32

receptor assay

Question 33

If the binding | reagent is a transport protein, it is a _______

Answer 33

Competitive protein-binding assay

Question 34

the first chemiluminescent label used in immunoassays, is a cyclic diacylhydrazide that emits light energy under alkaline conditions in the presence of peroxide and peroxidase.

Answer 34

Luminol

Question 35

compounds that absorb radiant energy of one wavelength and emit radiant energy of a longer wavelength in less than 10 −4 seconds.

Answer 35

Fluorescent labels (fluorochromes or fluorophores)

Question 36

atoms with unstable | nuclei that spontaneously emit radiation are radioactive and referred to as:

Answer 36

Radionuclides

Question 37

Radiolabeled antigen is also called:

Answer 37

tracer

Question 38

The earliest immunoassay was a ________ in which the radiolabeled Ag competed with unlabeled Ag for a limited number of binding sites (Ab)

Answer 38

Competitive immunoassay

Question 39

use a labeled reagent Ab to detect the Ag. Excess labeled Ab is required to ensure that the labeled Ab reagent does not limit the reaction.

Answer 39

Noncompetitive immunoassays/Immunometric assays

Question 40

physical separation is necessary and is | achieved by adsorption, precipitation, or interaction with a solid phase

Answer 40

Heterogeneous assays

Question 41

the activity or expression of the label depends on whether the labeled reactant is free or bound.

Answer 41

Homogeneous assays

Question 42

______ is porous and readily combines with small molecules to remove them from solution; ______ prevents nonspecific protein binding to the charcoal.

Answer 42

Charcoal; dextran

Question 43

use particles to trap small Ags, labeled or unlabeled.

Answer 43

Adsorption techniques

Question 44

occurs when the environment is altered, affecting the | solubility of protein.

Answer 44

Nonimmune precipitation

Question 45

Soluble Ag–Ab complexes can be precipitated by a second Ab that recognizes the primary Ab in the soluble complex. The result is a larger complex that becomes insoluble and precipitates.

Answer 45

Immune precipitation/Double Ab/2nd Ab method

Question 46

immobilize reagent Ab or Ag provides a method to | separate free from bound labeled reactant after washing.

Answer 46

Use of a solid phase

Question 47

Advantage of sandwich-type immunoassays

Answer 47

production of linear | calibration curves

Question 48

Disadvantage of sandwich-type immunoassays

Answer 48

Subjected to false (+) and false (-) interferences

Question 49

an engineered antibody whereby the antigen recognition site (Fab) originates from the mouse, while the constant portion contain sequences that are from the human species.

Answer 49

Chimeric antibody

Question 50

contains small segments from the mouse but sufficient in number to recognize the foreign antigen.

Answer 50

Humanized antibody

Question 51

a homogeneous competitive immunoassay in which low molecular weight Hps bound to particles compete with unlabeled analyte for the specific Ab.

Answer 51

Particle-enhanced turbidimetric inhibition immunoassay

Question 52

the unlabeled Ag in the sample competes with the labeled Ag for the Ab-binding sites; as the concentration of unlabeled Ag increases, less enzyme-labeled Ag can bind to the Ab. Therefore, more labeled Ag is free, and the enzymatic activity is greater.

Answer 52

``` enzyme-multiplied immunoassay technique (EMIT), ```

Question 53

are competitive, | homogeneous assays in which the genetically engineered label is β-galactosidase

Answer 53

Cloned enzyme donor immunoassays (CEDIAs)

Question 54

is an automated assay available on the AxSYM Analyzer (Abbott Laboratories). The microparticles serve as the solid phase, and a glass fiber matrix separates the bound labeled reagent.

Answer 54

Microparticle capture enzyme immunoassay (MEIA)

Question 55

fluid-phase unlabeled Ag is captured by Ab on the solid phase; after washing, the detector Ab (with a fluorescent label attached) reacts with the solid-phase captured Ag.

Answer 55

Solid-phase fluorescence immunoassays

Question 56

a heterogeneous, competitive immunoassay in which particles are used to localize the reaction and concentrate the fluorescence.

Answer 56

Particle concentration fluorescence immunoassay

Question 57

Fluorescein-labeled Ag and unlabeled Ag compete for rhodamine- labeled Ab. More unlabeled Ag lessens the amount of fluorescein-labeled Ag that binds; therefore, more fluorescence is present

Answer 57

Fluorescence excitation transfer immunoassay

Question 58

This homogeneous immunoassay uses polarized light to excite the fluorescent label. Polarized light is created when light passes through special filters and consists of parallel light waves oriented in one plane.

Answer 58

Fluorescence polarization immunoassay (FPIA)

Question 59

is an automated system (Thermo Fisher Scientific) that measures time-delayed fluorescence from the label europium.

Answer 59

Dissociation-enhanced lanthanide fluoroimmunoassay (DELFIA)

Question 60

heterogeneous, competitive assay with a tracer. 1 When bound tracer is measured, the signal from the label (CPM) is inversely related to the concentration of the unlabeled Ag in the sample.

Answer 60

RIA

Question 61

When used to microscopically identify bacteria or constituents in tissue (e.g., immune complexes deposited in vivo), the method is called

Answer 61

direct immunofluorescence (DIF) or direct fluorescence assay (DFA).

Question 62

The substrate is placed on a microscopic slide, serum is overlaid and allowed to react with the Ag, and the bound Ab is detected by the labeled antihuman globulin reagent.

Answer 62

indirect | immunofluorescence (IIF) or an indirect immunofluorescence assay (IFA).

Question 63

is used to classify cell lineage and identify the stage of cell maturation.

Answer 63

immunophenotyping

Question 64

is based on cells transported under fluidic pressure passing one by one through a laser beam.

Answer 64

Flow | cytometry

Question 65

is related to the size of the cell

Answer 65

Forward light scatter

Question 66

related to the | granularity of the cell.

Answer 66

Side light scatter

Question 67

are oligonucleotides that have been designed and engineered to bind to specific analytes and offer an alternative to Abs used in immunoassays.

Answer 67

Aptamers