Virology
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Presentation Transcript
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 • 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 • Antigen capture ELISA - eg rotavirus, HBsAg, HIV • Latex particle agglutination (LPA) eg rotavirus • Immunoflouresence - eg RSV, influenza,
Virus antigen detection: Latex particle agglutination for rotavirus
Virus antigen detection: Immunofluorescence of RSV-infected nasopharyngeal cells
Detection of viral genome • Genome amplification eg. PCR (polymerase chain reaction)
Detection of serological response • IgM detection • Detection of rising titre: ELISA, complement fixation test, haemagglutination inhibition
Detection of serological response: Antigen capture ELISA for HIV antibody
Case 1 • 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 1 • What additional tests would you request? • What your treatment options
Case 2 • 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 2 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 2 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 2 • Do you agree with the comments? • What additional test(s) would you request for after a discussion with the microbiologist?
Case 3 • 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 3 • Patient 1: Hepatits B surface antibody: >1000 mIU/ml • Patient 2: Hepatitis B surface antibody: <10 mIU/ml
Case 4 • 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 4 • Comment on the above report. • What additional tests would you request?
Case 4 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 • 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 5 • 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?