Immunological Assay Methods
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Questions and Answers

Why is it essential to collect an adequate sample for diagnosis?

  • To ensure the plant pathogen has survived on dead tissue
  • To prevent contamination by secondary microorganisms (correct)
  • To rule out soil pH or nutrients as contributors to the problem
  • To collect the whole plant, even if it's not possible
  • What is the primary reason for avoiding dead plants as samples?

  • The plant pathogen may not have survived on dead tissue (correct)
  • The plant tissue is difficult to sterilize
  • The plant tissue is too large to collect
  • The plant part is overrun with saprophytic microorganisms
  • What is the purpose of surface-sterilization in plant diagnosis?

  • To determine the plant's pH level
  • To analyze the plant's nutrient content
  • To detect plant pathogens in the soil
  • To eliminate saprophytic microorganisms from the plant tissue (correct)
  • What is the recommended method for surface-sterilization?

    <p>Soaking the tissue in a 10-20% bleach solution</p> Signup and view all the answers

    When is it necessary to collect whole plants for diagnosis?

    <p>When there is suspicion of root problems contributing to foliar symptoms</p> Signup and view all the answers

    Why are soil samples sometimes collected around damaged and healthy plants?

    <p>To understand the role of soil pH or nutrients in the plant problem</p> Signup and view all the answers

    What is the primary concern when collecting samples from trees?

    <p>Collecting the whole plant is not possible</p> Signup and view all the answers

    What is the ideal size of a turf grass sample for diagnosis?

    <p>A 7&quot; × 7&quot; area with sufficient depth to include the roots</p> Signup and view all the answers

    Why is it essential to collect a representative sample from symptomatic or suspected parts of a plant?

    <p>To ensure the sample is representative of the whole plant</p> Signup and view all the answers

    What is the primary advantage of collecting an adequate sample for diagnosis?

    <p>It ensures accurate detection of plant pathogens</p> Signup and view all the answers

    Study Notes

    Pathogen Detection and Identification

    • Common methods used for pathogen detection and identification include ELISA, Western blot, Immunofluorescence microscopy, and Rapid Immunodiagnostic Tests.
    • Biochemical methods employed include Enzyme Assay, Protein/Metabolite Profiling, RNA and Gene Expression Analysis.
    • Molecular analytical methods used include PCR-based detection assays, Real-time Quantitative PCR (qPCR), Polymerase Chain Reaction (PCR), Loop-mediated Isothermal Amplification (LAMP), and Next-generation Sequencing (NGS).

    Pathogenicity Test

    • Pathogenicity test is the ultimate test to demonstrate proof of pathogenicity.
    • The test involves reproducing the damage using the suspected pathogen.
    • Sterilization of plant material is necessary before testing, usually done with ethanol, peroxide, or n-alkyl products.
    • Exposure time to the sterilizing agent ranges from a few seconds to several minutes, depending on the structure of the plant tissue.

    Pathogen Culture

    • About 85% of plant diseases are caused by fungi.
    • Sterile tissue samples are sectioned and placed on sterile media culture (petri dishes).
    • Culture is maintained at room temperature or in an incubator at a specific temperature (e.g., 28°C–30°C for bacterial growth or ~25°C for fungal growth).
    • The type of media culture is important for the type of microorganism to be isolated.

    Serological Methods

    • Serology is the study of immunological reactions between antigens and antibodies.
    • Adequate sample collection is necessary for a proper diagnosis, and the sample size varies depending on the type of problem and plant.

    Sample Collection and Preparation

    • Avoid dead plants as samples, as they may not have surviving pathogens or may be overrun with saprophytic microorganisms.
    • Collect representative samples from symptomatic or suspected parts, or whole plants if possible.
    • Soil samples may be necessary when soil pH or nutrients are suspected to be contributing to the plant problem.

    Isolation and Recovery

    • Surface-sterilization is necessary to reduce surface contaminants and saprophytes.
    • The most common method is soaking the tissue in a diluted bleach solution (e.g., 10-20% or 0.5–1% NaOCl).

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    Description

    This quiz covers various immunological assay methods, including ELISA, Western blot, Immunofluorescence microscopy, and Rapid Immunodiagnostic Tests, used to identify unknown bacteria.

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