Survey of hepatites VirusRelationshipTransmission HAVPicornavirusfaecal-oral HBVSpecial group of DNA viruses sexual, blood HCV (+ HGV)Flavivirusblood HDVDelta agens – viroid sexual, blood HEVRelated 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.

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

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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 D = 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