Survey of hepatitesVirus Relationship Transmission

HAV Picornavirus faecal-oral

HBV Special group of DNA viruses

sexual, blood

HCV (+ HGV)

Flavivirus blood

HDV Delta agens – viroid

sexual, blood

HEV Related to caliciviruses

faecal-oral

Hepatites• liver inflamations, with jaundice as a

typical symptoma. Patient has fever, intestinal problems, yellow colour of sclera or skin, changed urine and stool colour etc. Hepatitis B, C and D may become chronical, and they may act as precanceroses.

Patients with jaundice

medicine.ucsd.edu/Clinicalimg/skin-jaundice.html.

http://www.gihealth.com/images/imgJaundiceBig.jpg

Hepatitis A (HAV)

• No chronicity• Dg.: ELISA method, we determine anti-

HAV IgM and total Ig• Vaccination is available

Hepatitis E • HEV. IgM and IgG ELISA detection, PCR

is in developement• No chronicity• ! pregnant ladies – fulminant status

www.pegasys.com/hcp/efficacy-hepatitis-b.aspx

HBV• Hepatitis C may become chronical,

and may act as precancerosis • Treatment: interferons,

hepatoprotective drugs, other symptomatic therapy

• Vaccination agains hepatitis B is part of normal vaccination calendar in many countries

HBV diagnostics• 2 core antigens HBcAg and HBeAg are

present, 1 surface antigen HBsAg. Only 2 of them can be found in patient‘s serum: HBsAg and HBeAg. HBsAg is suitable for screening

• Antibodies may be assessed against all three of them: anti-HBs, anti-HBe and anti-HBc.

• Also PCR is sometimes used, hepatal enzymes detection etc.

• Interpretation needs combination of results

Diagnostics of hepatitis C (HCV)• ELISA detection of IgM and IgG, and

also PCR method.• Hepatitis C may become chronical,

and may act as precancerosis• treatment by interferons is used• hepatoprotective drugs and

other symptomatic therapy are used

Delta agens (HDV)

• is a viroid particle• may infect humans together with HBV

(coinfection), or following after such infection (superinfection)

• Presence of delta agens worsens prognosis of viral hepatitis

• Determining delta antigen (HDAg), antibodies (anti-HD) or viral RNA PCR

HIV – Human Immunodeficiency Virus

• Retrovirus, it has reverse transcriptase (an enzyme transcripting RNA into DNA)

• Transmitted by blood, sexually and moter-to-child transmission

• There is a lot of drugs against HIV, but ther effectivity is limited

Diseases caused by HIV

•Virus attacs cellullar immunity•1. After a non-specific primary infection there is a long latent period

•2. Generalized lymphadenopaty occur•3. complete AIDS. The patients have symptomas of opportune infections (toxoplasmosis, pneumocystosis, various mycoses etc.) and tumors

HIV diagnostics

• Antibodies. When the result looks like positive, the serum specimen is sent to reference laboratory, that confirms the result using western blotting

• Direct diagnostics: PCR can be performed. Virus isolation is possible, but very difficult

Practical notes to tasks• Unfortunatelly, the patients of individual

parts of your tasks are not related, except tasks 1a) and 1b). So do not try to perform „final conclusion“ of tasks 2a) + 2b) + 2c) + 2d), it is not possible

• In ELISA reactions, write the „coordinates“ of your patients to the protocol (e. g. A2, C3, E5 etc.)

• In task 1 b), please, write negative patients, not positive ones (they would be too many)

• Picture in 3a) draw rather black on white background, not white on black backround

Task 1 – diagnostics of HAV• Task 1a) is determination of anti-HAV IgM.

Counting cut off: average C1 / D1• Task 1b) is determination of total anti-HAV

antibodies (IgM + IgG + all other classes)• Counting cut off (1a + b):

– Average of „c. o.“ wells C1 and D1 = cut off– Values 110 % of cut off and more = positive– Values 90 % of cut off and less = negative– Values 90–110 % of cut off = borderline

• Assessment of IgM + total antibodies is an alternative to assessment of IgM a IgG‘s

Task 2: Determination of HBV markers

• 2a + b) Determination of HBsAg and HBeAg

• 2c + d) Determination of anti-HBs, anti-HBe– Average of „c. o.“ wells C1 and D1 = cut off– Values 110 % of cut off and more = positive– Values 90 % of cut off and less = negative– Values 90 – 110 % of cut off = borderline

Task 3: HCV diagnostics• 3a): PCR, electroforesis• 3b): anti-HCV, ELISA method• Average of „negative control“ wells

B1, C1 and D1 + 0.050 = cut off• Values 110 % of cut off and more =

positive• Values 90 % of cut off and less =

negative• Values 90–110 % of cut off =

borderline

Task 4: ELISA anti-HIV

• Average of „c. o.“ wells C1 and D1 = cut off

• Values 110 % of cut off and more = positive• Values 90 % of cut off and less = negative• Values 90 – 100 % of cut off = borderline