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Virology. 22-2-11. Diagnostic methods in virology. Detection of virus Detection of viral antigen Detection of viral genome Detection of serological response. Detection of viral antigens. Antisera (mono or polyclonal) are used to detect viral antigen

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Virology

Virology

22-2-11


Diagnostic methods in virology
Diagnostic methods in virology

  • Detection of virus

  • Detection of viral antigen

  • Detection of viral genome

  • Detection of serological response


Detection of viral antigens
Detection of viral antigens

  • Antisera (mono or polyclonal) are used to detect viral antigen

  • These assays can be sensitive as they can detect antigens of disrupted viral particles

  • They are pathogen specific - separate immuno-assay must be used for each pathogen

  • In contrast electron microscopy and culture are catch all techniques which detect all pathogens


Methods
Methods

  • Antigen capture ELISA - eg rotavirus, HBsAg, HIV

  • Latex particle agglutination (LPA) eg rotavirus

  • Immunoflouresence - eg RSV, influenza,







Detection of viral genome
Detection of viral genome nasopharyngeal cells

  • Genome amplification eg. PCR (polymerase chain reaction)


Viral genome detection qpcr
Viral genome detection: qPCR nasopharyngeal cells


Viral genome detection pcr
Viral genome detection: PCR nasopharyngeal cells


Detection of serological response
Detection of serological response nasopharyngeal cells

  • IgM detection

  • Detection of rising titre: ELISA, complement fixation test, haemagglutination inhibition



Case 1
Case 1 HIV antibody

  • The following is a report from a lady with a 10-year history of fatigue, anorexia and elevated LFTs.

    Hepatitis B surface antigen: negative

    Hepatitis C antibody: positive

    Hepatitis C antibody immunoblot confirmation: positive Banding pattern: c100 c33 c22 NS5 3+ 3+ 4+ 4+


Case 11
Case 1 HIV antibody

  • What additional tests would you request?

  • What your treatment options


Case 2
Case 2 HIV antibody

  • You are given the following results on two different patients.

  • Are there any differences between the profiles?

  • What implications (if any) are there for these differences?


Case 21
Case 2 HIV antibody

Patient 1:

TestsResults

EIA for HBsAg Positive

***Infection Risk***

Hepatitis B e antigen Negative

Hepatitis B e antibody Positive

Hepatitis B core IgM antibody Negative

Report comment: Hepatitis B result profile consistent with

chronic hepatitis B infection.


Case 22
Case 2 HIV antibody

Patient 2:

TestsResults

EIA for HBsAg Positive

***Infection Risk***

Hepatitis B e antigen Positive

***High Infection Risk***

Hepatitis B e antibody Negative

Hepatitis B core IgM antibody Positive: 250 IU/ml.

Report comment: Hepatitis B result profile consistent with

chronic hepatitis B infection.


Case 23
Case 2 HIV antibody

  • Do you agree with the comments?

  • What additional test(s) would you request for after a discussion with the microbiologist?


Case 3
Case 3 HIV antibody

  • You have vaccinated 2 patients against hepatitis B virus for the purpose of foreign travel.

  • What test would you request to assess the vaccination status?

  • Comment on the following results.


Case 31
Case 3 HIV antibody

  • Patient 1:

    Hepatits B surface antibody: >1000 mIU/ml

  • Patient 2:

    Hepatitis B surface antibody: <10 mIU/ml


Case 4
Case 4 HIV antibody

  • A patient presents with malaise, elevated LFTs and a history of intravenous drug abuse. You are given the following result:

    Hepatitis B surface antigen: Positive

    Hepatitis B surface antibody: Negative

    Report comment: If this is the first positive serological test on this patient, please repeat to confirm.


Case 41
Case 4 HIV antibody

  • Comment on the above report.

  • What additional tests would you request?


Case 42
Case 4 HIV antibody

Hepatitis B surface antigen: Positive

***Infection Risk***

Hepatitis B e antigen: Positive

***High Infection Risk***

Hepatitis B e antibody: Negative

Hepatitis B core IgM antibody: Negative

Hepatitis B core total antibody: Positive

Report comment: ………..

  • What is your report comment?


Case 5
Case 5 HIV antibody

  • A 23 year old female patient presents to the A/E department with malaise, lymphadenopathy, malaise and a rash. She has a history of unsafe sexual activity.

  • What investigations would you request?


Case 51
Case 5 HIV antibody

  • Shown below is one of the patient’s laboratory results:

    TestsResults

    EIA for anti-HIV 1 (Organon) Positive

    EIA for anti-HIV 1+2 (Murex) Positive

    Report comment: ***confirmed anti-HIV 1 positive***

  • What additional tests would you require?

  • What would you advise the patient?


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