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    Swu Phinma: Dorothy Nacilla

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    Zoe Gellai
    PCR is a method used to detect the presence of viral RNA through amplification of genetic material. It works by converting RNA to DNA using reverse transcriptase, then replicating the DNA using the polymerase chain reaction. This allows detection of even small amounts of viral genetic material in samples taken from the nose and throat. A fluorescent probe is used to detect the amount of viral DNA present, with a lower cycle threshold indicating a higher viral load in the original sample. PCR testing is useful for diagnosing current SARS-CoV-2 infection by detecting the virus's genetic material in a patient's body fluids or tissues.

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    Swu Phinma: Dorothy Nacilla

    Uploaded by

    Zoe Gellai
    26 views17 pages
    PCR is a method used to detect the presence of viral RNA through amplification of genetic material. It works by converting RNA to DNA using reverse transcriptase, then replicating the DNA using the polymerase chain reaction. This allows detection of even small amounts of viral genetic material in samples taken from the nose and throat. A fluorescent probe is used to detect the amount of viral DNA present, with a lower cycle threshold indicating a higher viral load in the original sample. PCR testing is useful for diagnosing current SARS-CoV-2 infection by detecting the virus's genetic material in a patient's body fluids or tissues.

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    PCR is a method used to detect the presence of viral RNA through amplification of genetic material. It works by converting RNA to DNA using reverse transcriptase, then replicating the DNA using the polymerase chain reaction. This allows detection of even small amounts of viral genetic material in samples taken from the nose and throat. A fluorescent probe is used to detect the amount of viral DNA present, with a lower cycle threshold indicating a higher viral load in the original sample. PCR testing is useful for diagnosing current SARS-CoV-2 infection by detecting the virus's genetic material in a patient's body fluids or tissues.

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    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    26 views17 pages

    Swu Phinma: Dorothy Nacilla

    Uploaded by

    Zoe Gellai
    PCR is a method used to detect the presence of viral RNA through amplification of genetic material. It works by converting RNA to DNA using reverse transcriptase, then replicating the DNA using the polymerase chain reaction. This allows detection of even small amounts of viral genetic material in samples taken from the nose and throat. A fluorescent probe is used to detect the amount of viral DNA present, with a lower cycle threshold indicating a higher viral load in the original sample. PCR testing is useful for diagnosing current SARS-CoV-2 infection by detecting the virus's genetic material in a patient's body fluids or tissues.

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    SWU PHINMA

    Dorothy Nacilla
    PCR Testing in
    SARS-CoV2
    Introduction to PCR

    Polymerase
    Chain Reaction
    Testing the presence of viral RNA using real time Reverse
    Transcriptase Polymerase Chain Reaction or RT-PCR.

    SARS - COV2
    real time
    RT-PCR
    PCR is a test
    to
    determine
    genetic
    material
    from specific
    organism
    such as virus.
    Specimen for SARS-CoV2:
    Oropharyngeal and
    Nasopharyngeal
    Others: Saliva

    Extract all RNA in the


    sample but RNA is more
    difficult to work with in
    the lab than DNA.
    Overview of real time RT-PCR
    DNA can be easily amplified & quantified using simple,
    accurate method called PCR or polymerase chain reaction.
    PCR does not work with RNA.So,we convert RNA to DNA
    using an enzyme called Reverse transcriptase.

    Resulting DNA: Complementary DNA ( cDNA )


    Sequence is complementary to that of the original RNA
    strand.
    Note: created a collection of cDNA's representative of all
    the RNA present in the original sample

    RNA : includes virus,bacteria,patient's own cell


    A way to determine presence of SARS-COV2 cDNA : PCR
    Invented: 1985 by Kary Mullis in which a small amount of DNA can be
    copied in large quantities over a short period of time.

    Amplify + detect specific DNA molecule = viral cDNA

    3 STEPS: 40x
    1. Denaturation : 2 strands separated
    2. Annealing: a pair of primers* specific to SARS-CoV2 cDNA
    attaches/anneals to each DNA strands
    3. Elongation: enzyme polymerase adds nucleotides to the ends of the
    primers using the original DNA strand as a template
    *Primers are short nucleotide sequences complementary to unique
    sequence in the viral cDNA
    Specificity of primers ensures that they only bind to viral
    cDNA and not to other cDNA present in the sample.

    1 double stranded DNA to 2


    These 3 steps are repeated with the amount of DNA doubling in every cycle.
    In real time pcr, a probe
    is added during the pcr
    process which gives off
    fluorescence whenever
    a new DNA molecule is
    formed.
    Increase in viral cDNA = increase flourescence signal
    When the level of fluorescence exceeds a certain
    threshold,the signal is significantly above the background
    level.
    CT= cycle threshold, is the number of cycles required for
    the fluorescent signal to exceed that specific threshold
    value.

    Inversely proportional: The lower the CT value,the more


    RNA present in the original sample indicating higher viral
    load.
    For SARS-CoV2
    RESULTS +
    CLINICAL
    CORRELATION
    Thank You!

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