Bead-Based Immunoassay Techniques Quiz
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Questions and Answers

Which concentration unit is used for the analyte GH (baseline)?

  • Millimolar
  • Micromolar (correct)
  • Picomolar
  • Nanomolar
  • Which detection technology is used in the Vitros 250 analyzer?

  • Fluorimetry
  • Luminometry
  • Dry Chemistry (correct)
  • Slide Technology
  • Which company offers the Cobas 8000 analyzer?

  • Abbott
  • Orthoclinical Diagnostics
  • Roche (correct)
  • Siemens
  • Which type of assay involves phase separation and separation of analytes bound to solid phase from that in solution?

    <p>Heterogenous assays</p> Signup and view all the answers

    Which of the following factors can affect the performance of labs in the RCPA EQA scheme?

    <p>The inherent characteristics of the method</p> Signup and view all the answers

    What type of bias is apparent in the lab's results compared to the median results from other labs?

    <p>Negative proportional bias</p> Signup and view all the answers

    What type of distribution does the data for all labs show and why?

    <p>Bimodal distribution due to inherent characteristics of the method</p> Signup and view all the answers

    What is the analytical principle of the assay specified by the lab for measurement of prolactin?

    <p>Immunoassays</p> Signup and view all the answers

    Which type of assay uses an excess of antibodies and is typically used for higher molecular weight antigens?

    <p>Sandwich Assay (ELISA Example)</p> Signup and view all the answers

    Which one of the following best describes the allowable limits of performance?

    <p>The acceptable range of values for a specific measurement.</p> Signup and view all the answers

    Which type of assay involves a fixed amount of enzyme-labeled antigen competing with antigen from the patient sample for binding to a limited amount of antibody?

    <p>Competitive ELISA</p> Signup and view all the answers

    If a laboratory can measure plasma sodium with a standard deviation (SD) of 3.0 mmol/L, is that considered satisfactory?

    <p>No, it is not considered satisfactory.</p> Signup and view all the answers

    Which type of assay offers higher sensitivity compared to measuring absorbance and involves measuring either a fluorescence signal or chemiluminescence?

    <p>Chemiluminescent ELISA</p> Signup and view all the answers

    What type of auto-analyzer would be required to measure a steroid present at a concentration of 12 pmol/L?

    <p>A dedicated immunoassay analyzer.</p> Signup and view all the answers

    Which type of assay uses luminescent labels that emit a photon of light and offers highly sensitive detection with no background light?

    <p>Ruthenium complexes</p> Signup and view all the answers

    Which type of assay is the Abbott Architect Immunoassay (Chemiflex system)?

    <p>A heterogenous assay</p> Signup and view all the answers

    What is the purpose of the wash steps in heterogenous assays?

    <p>To remove unreacted or unbound fraction</p> Signup and view all the answers

    Which type of assay is the Roche Immunochemiluminometric assay (ELECSYS system)?

    <p>A competitive or titrimetric assay</p> Signup and view all the answers

    What is the purpose of the streptavidin coated magnetized beads in the Roche Immunochemiluminometric assay (ELECSYS system)?

    <p>To draw magnetized beads to the electrode</p> Signup and view all the answers

    According to the text, what is the principle difference between two-step assays and one-step assays in immunoassays?

    <p>Two-step assays are prone to the hook effect, while one-step assays are not.</p> Signup and view all the answers

    According to the text, what is the purpose of a proficiency program in laboratory testing?

    <p>To compare the performance of different laboratories</p> Signup and view all the answers

    According to the text, what type of auto-analyzer is required to measure a steroid at a concentration of 12 pmol/L?

    <p>A mass spectrometer</p> Signup and view all the answers

    According to the text, what are the two major types of auto-analyzers in a clinical biochemistry laboratory?

    <p>Chemiluminescence analyzers and mass spectrometers</p> Signup and view all the answers

    Which term describes assays that can be done entirely in the liquid phase, with no requirement to separate any component of the reagents to get a readout?

    <p>Homogeneous</p> Signup and view all the answers

    What are the similarities and differences between Siemens Immulite and Roche Elecsys immunoassays?

    <p>Both use chemiluminescence as a detection method, but Elecsys is more sensitive</p> Signup and view all the answers

    What are the two major sources of interference that the Roche Elecsys system is subject to, which are not seen in most other sensitive immunometric assays?

    <p>Antibodies and enzymes</p> Signup and view all the answers

    What is the difference between chemiluminescence and fluorescence, and why do immunoassays using chemiluminescence yield a higher sensitivity?

    <p>Chemiluminescence is the emission of light from a chemical reaction, while fluorescence is the emission of light from an excited state. Immunoassays using chemiluminescence yield a higher sensitivity due to the higher intensity of light emitted in chemiluminescent reactions.</p> Signup and view all the answers

    Study Notes

    Concentration Units and Detection Technologies

    • The analyte GH (baseline) is measured in concentration units of μIU/mL.
    • The Vitros 250 analyzer uses chemiluminescence detection technology.
    • The Cobas 8000 analyzer is offered by Roche.

    Assay Types

    • Immunosorbent assays (ISAs) involve phase separation and separation of analytes bound to solid phase from that in solution.
    • Excess reagent immunoassays use an excess of antibodies and are typically used for higher molecular weight antigens.
    • Competitive assays involve a fixed amount of enzyme-labeled antigen competing with antigen from the patient sample for binding to a limited amount of antibody.

    Performance and Bias

    • Factors that can affect the performance of labs in the RCPA EQA scheme include instrumental, procedural, and reagent variations.
    • A positive bias is apparent in the lab's results compared to the median results from other labs, indicating a systematic error.
    • The data for all labs shows a non-normal (skewed) distribution, indicating variability in lab performance.

    Analytical Principles and Assay Characteristics

    • The analytical principle of the assay specified by the lab for measurement of prolactin is immunometric.
    • Chemiluminescence assays offer higher sensitivity compared to measuring absorbance and involve measuring either a fluorescence signal or chemiluminescence.
    • Chemiluminescence assays use luminescent labels that emit a photon of light and offer highly sensitive detection with no background light.

    Assay Systems and Instruments

    • The Abbott Architect Immunoassay (Chemiflex system) is a type of heterogeneous assay.
    • The Roche Immunochemiluminometric assay (ELECSYS system) is a type of heterogeneous assay.
    • The wash steps in heterogeneous assays are necessary to remove excess reagents and minimize non-specific binding.
    • The streptavidin coated magnetized beads in the Roche Immunochemiluminometric assay (ELECSYS system) are used to capture the analyte-antibody complex.

    Proficiency Programs and Auto-analyzers

    • The purpose of a proficiency program in laboratory testing is to ensure accuracy and reliability of laboratory results.
    • A proficiency program evaluates laboratory performance by comparing results with peer labs.
    • To measure a steroid at a concentration of 12 pmol/L, a high-sensitivity auto-analyzer is required.
    • The two major types of auto-analyzers in a clinical biochemistry laboratory are random-access and batch analyzers.

    Immunoassay Characteristics

    • Homogeneous assays are those that can be done entirely in the liquid phase, with no requirement to separate any component of the reagents to get a readout.
    • Two-step assays involve a separate incubation and washing step, whereas one-step assays do not require a separate washing step.
    • The principle difference between two-step assays and one-step assays is the separation of incubation and washing steps.

    Interference and Assay Comparison

    • The Roche Elecsys system is subject to interference from heterophilic antibodies and rf-F(ab')2, which are not seen in most other sensitive immunometric assays.
    • Siemens Immulite and Roche Elecsys immunoassays share similarities in their use of chemiluminescence detection, but differ in their assay design and reagent composition.
    • Chemiluminescence assays yield a higher sensitivity than fluorescence assays due to the ability to detect single photon events.

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    Description

    Test your knowledge on bead-based immunoassay techniques with this quiz! Learn about the different assay characteristics, including sandwich assays, chemiluminescent reporter systems, and enzyme-linked immunoassays. Challenge yourself and expand your understanding of immunoassay methods.

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