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    Laboratory Diagnosis of Viral Infection

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    studymedic
    This document discusses laboratory diagnosis of viral infections through direct detection, isolation, and serology. Direct detection can identify virus particles, antigens, or nucleic acids using electron microscopy, immunofluorescence, ELISA, PCR, or probe hybridization. Viruses can be isolated from clinical specimens by culturing them in tissue culture, chick embryos, or laboratory animals. Identification involves observing cytopathic effects or using neutralization tests. Serology detects antibodies using immunofluorescence, ELISA, hemagglutination inhibition, neutralization tests, or complement fixation. Together, these methods allow identification of viruses through direct, culture-based, and immunological approaches.

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    Laboratory Diagnosis of Viral Infection

    Uploaded by

    studymedic
    315 views37 pages
    This document discusses laboratory diagnosis of viral infections through direct detection, isolation, and serology. Direct detection can identify virus particles, antigens, or nucleic acids using electron microscopy, immunofluorescence, ELISA, PCR, or probe hybridization. Viruses can be isolated from clinical specimens by culturing them in tissue culture, chick embryos, or laboratory animals. Identification involves observing cytopathic effects or using neutralization tests. Serology detects antibodies using immunofluorescence, ELISA, hemagglutination inhibition, neutralization tests, or complement fixation. Together, these methods allow identification of viruses through direct, culture-based, and immunological approaches.

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    This document discusses laboratory diagnosis of viral infections through direct detection, isolation, and serology. Direct detection can identify virus particles, antigens, or nucleic acids using electron microscopy, immunofluorescence, ELISA, PCR, or probe hybridization. Viruses can be isolated from clinical specimens by culturing them in tissue culture, chick embryos, or laboratory animals. Identification involves observing cytopathic effects or using neutralization tests. Serology detects antibodies using immunofluorescence, ELISA, hemagglutination inhibition, neutralization tests, or complement fixation. Together, these methods allow identification of viruses through direct, culture-based, and immunological approaches.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PPT, PDF, TXT or read online from Scribd
    Download as ppt, pdf, or txt
    315 views37 pages

    Laboratory Diagnosis of Viral Infection

    Uploaded by

    studymedic
    This document discusses laboratory diagnosis of viral infections through direct detection, isolation, and serology. Direct detection can identify virus particles, antigens, or nucleic acids using electron microscopy, immunofluorescence, ELISA, PCR, or probe hybridization. Viruses can be isolated from clinical specimens by culturing them in tissue culture, chick embryos, or laboratory animals. Identification involves observing cytopathic effects or using neutralization tests. Serology detects antibodies using immunofluorescence, ELISA, hemagglutination inhibition, neutralization tests, or complement fixation. Together, these methods allow identification of viruses through direct, culture-based, and immunological approaches.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PPT, PDF, TXT or read online from Scribd
    Download as ppt, pdf, or txt
    of 37

    Laboratory

    Laboratory Diagnosis
    Diagnosis
    of
    of Viral
    Viral Infection
    Infection
    Detection – Isolation - Serology
    What is a virus?

    It is a segment of either RNA or


    DNA protected by a protein coat
    and in some families of viruses a host
    derived envelope with attached viral
    proteins
    It lacks:
    - Protein synthesizing machinery
    - Energy producing system
    - No mitochondria
    - No stores of amino acids, nucleotides energy
    rich molecules
    It is a compulsory intra cellular parasite
    It depends on three main
    principles:
    Direct detection Isolation on: Serology using:
    of:

    Virus particles • Tissue culture IF

    Viral antigen • Chick embryo HI

    Viral nucleic acid • Laboratory NT


    animals

    Cytopathology ELISA
    I. Direct detection of virus
    particle
    Direct detection of virus:
    • particle,
    • viral antigen,
    • or viral nucleic acid in clinical
    specimens
    I. Direct detection of virus
    particle
    Direct detection could be done by one of the following:
    •
    Particle
    • Electron microscopy.
    –
    Antigen detection
    •
    Fluorescent antibody test.
    ELISA
    Immunodiffusion.
    Nucleic acid
    •

    1- 5- PCR.
    1. EM detection of
    corona virus
    Hepatitis B virus
    EM picture of rabies
    virus
    2. Detection of virus by
    Immunofluorescent Technique

    Diagramatic presentation of
    IF technique
    2. Detection of virus by
    Immunofluorescent Technique
    IF staining of rabies
    infected brain cells
    3. ElISA
    4. Immune diffusion
    5. Nucleic acid
    techniques
    • PCR

    • Probe Hybridization
    II- Isolation and
    identification
    II- Isolation and identification of the virus
    from clinical specimens: three main
    systems are used for viral isolation:
    1- Tissue culture.

    2- Chick embryo.

    3- Laboratory animals
    Tissue culture preparation:

    From the desired tissue the following steps are followed:

    • Mince into 1mm fragments.

    • Incubate with proteolytic enzyme (trypsin) to disperse


    the cells.

    • Add growth media to make a cell suspension.

    • Incubate in stationary flasks or tubes, cells settle on


    the dependent surface and grow into confluent
    monolayer.

    • Re-disperse monolayer cells and increase number of


    cultures for cell culture passage.
    Virus isolation in tissue
    culture cell line
    Viral identification:

    This is achieved by:


    (a) The effect on cell culture:
    i.e. cytopathic effect,

    (b) Neutralization test. This is based on the


    neutralization of the virus infectivity by
    mixing it with specific antibody before
    inoculation into cultures.
    Cytopathic effect
    Neutralization Test
    Following virus isolation: 1.Divide culture yield into small volume in a set of
    test tubes
    2. Prepare the panel of antisera against which the virus isolate is to be
    challenged
    2. To each test tube add one antisera and leave one as a virus control and
    one as serum control
    1 2 3

    B
    • Incubate for one hour then inoculate
    each into cell culture tubes, incubate
    and observe daily.
    1 2 3

    Principle of Neutralization test


    Rabid Virus

    Diagramatic presentation
    of rabies virus

    IF staining of rabies
    infected brain cells

    Negi bodies
    Isolation in embryonated
    hen’s eggs
    Inoculation into the
    amniotic cavity of the chick
    embryo.
    Inoculation into the yolk sac
    of the chick embryo
    A VIRUS INOCULATION BEING DROPPED
    ONTO THE CHORIOALLANTOIC
    MEMBRANE OF THIRTEEN DAY OLD
    CHICK EMBRYO .
    Herpes virus lesion on the
    chorioallantoic membrane
    Haemagglutination
    & Haemagglutination inhibition

    HAI

    HA
    III- Serological demonstration
    of the antibodies by:

    1- Immunofluorescence (IIF).
    2- Enzyme immunosorbant assay (ELISA).
    3- Haemagglutination inhibition test (HI).
    4- Neutralization test (NT).
    5- Complement fixation
    Serology
    Neutralization
    • Neutralization.

    Standardized antiserum is used


    HI
    • HI tests to detect specific
    antibodies to a virus in the patient’s
    serum
    Main material used
    • Standardized virus.
    THANK YOU

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