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    Molecular Biology Lab Finals

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    dokutah
    This document discusses specimen collection, storage, and transport for diagnosing viral diseases. It notes that viruses are most concentrated early in infection so samples should be collected soon after symptoms start. Samples like swabs, blood, urine and stool can be taken from infected sites and transported in viral transport medium, which protects viruses during refrigerated transport for up to 48 hours. Viruses are then isolated using cell cultures, including primary cells directly from tissues or established cell lines, and identified using electron microscopy, antibody detection, antigen detection or nucleic acid tests.

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    Molecular Biology Lab Finals

    Uploaded by

    dokutah
    16 views2 pages
    This document discusses specimen collection, storage, and transport for diagnosing viral diseases. It notes that viruses are most concentrated early in infection so samples should be collected soon after symptoms start. Samples like swabs, blood, urine and stool can be taken from infected sites and transported in viral transport medium, which protects viruses during refrigerated transport for up to 48 hours. Viruses are then isolated using cell cultures, including primary cells directly from tissues or established cell lines, and identified using electron microscopy, antibody detection, antigen detection or nucleic acid tests.

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    Molecular-Biology-Lab-Finals

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    This document discusses specimen collection, storage, and transport for diagnosing viral diseases. It notes that viruses are most concentrated early in infection so samples should be collected soon after symptoms start. Samples like swabs, blood, urine and stool can be taken from infected sites and transported in viral transport medium, which protects viruses during refrigerated transport for up to 48 hours. Viruses are then isolated using cell cultures, including primary cells directly from tissues or established cell lines, and identified using electron microscopy, antibody detection, antigen detection or nucleic acid tests.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    16 views2 pages

    Molecular Biology Lab Finals

    Uploaded by

    dokutah
    This document discusses specimen collection, storage, and transport for diagnosing viral diseases. It notes that viruses are most concentrated early in infection so samples should be collected soon after symptoms start. Samples like swabs, blood, urine and stool can be taken from infected sites and transported in viral transport medium, which protects viruses during refrigerated transport for up to 48 hours. Viruses are then isolated using cell cultures, including primary cells directly from tissues or established cell lines, and identified using electron microscopy, antibody detection, antigen detection or nucleic acid tests.

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    Application of Molecular Biology in brain biopsy and sometimes serum

    Medical Virology – Specimen Collection, are used.


    Storage and Transport
    d. Urogenital infections – needle
    aspirates and endocervical and
    urethral swabs
    Specimen Processing for Diagnosing
    Viral Diseases e. Gastrointestinal tract – stool
    samples and rectal swabs
     Viruses are in highest
    concentrations during the first f. Eye infections – eye swabs and
    several days following onset of corneal scrapings
    symptoms. Therefore, samples
    should be collected early in the
    disease course. Sample Transport
     Viruses are fastidious: sensitive to Samples for viral culture must be placed
    heat and drying into a Viral Transport Medium (VTM).
     Clinical specimens should be VTM Contains:
    handled with care and delivered to
    the laboratory as soon as possible a. Buffered Saline

     NEVER use Calcium Alginate b. Protein Stabilizers


    swabs: use DACRON or RAYON c. Antimicrobials that inhibit bacterial
    SAMPLES: and fungal growth

    1. Heparinized Whole Blood – for  Samples for viral cultures can be


    viral / bacterial culture refrigerated in VTM for about 48
    hours, they should never be frozen
    2. EDTA Plasma – for Serology and at -20°C. Samples can be stored at -
    PCR 70°C; however, infectivity will be
    diminished.
    Samples should be generally come from the
    infected site.
    a. Skin Infections – rash site and Viral Isolation
    depending on the virus, serum or
    urine
    b. Respiratory infections – sputum or 1. CELL CULTURE
    throat swabs  Important means for diagnosing viral
    c. Central Nervous System infections
    (meningitis and encephalitis) – for  Requires nutritionally rich complex
    diagnosis of meningitis, CSF and media
    serum, as well as stool or throat
    swabs, can be collected because  The media often contains fecal calf
    viruses are sometimes shed into serum as a nutrient
    these sites. In case of encephalitis,
     Some viruses have not yet been
    grown in vitro.
    very sensitive. For these reasons,
    electron microscopy is not
    2. PRIMARY CELL CULTURES
    commonly used.
     Derived directly from tissues
     Can only be maintained for a short
    time inside the laboratory
    Other methods of detection
     Example: Primary Monkey Kidney
    Cells 1. Detection of host antibodies directed
    against specific viruses
     Transferring cells from one container
    to another is called “splitting” or 2. Direct detection of viral antigens in
    “passaging” clinical specimens
    3. ESTABLISHED CELL LINES 3. Viral gene probes and nucleic acid
    amplification (e.g., polymerase chain
     Also referred to as low passage or
    reaction [PCR])
    finite cell lines
     Can be maintained longer than
    primary cell lines
     Examples: WI – 38, MRC – 5, IMR -
    90
    4. CONTINUOUS CELL LINES
     Are altered cells that can be
    maintained indefinitely
     Examples: HeLa Cells, HEp – 2,
    A549, and Vero Cells
     They can be used to grow HSV,
    VZV, CMV, Adenovirus and
    Rhinoviruses.

    Electron Microcopy
     Due to size, most individual virions
    can only be seen by electron
    microscopy.
     Electron Microscopy is sometimes
    used to identify Norwalk viruses,
    Astrovirus, Calicivirus, and
    Coronavirus.
     Electron Microscope is expensive,
    requires expertise and is usually

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