1. Viruses Only 1 NA Small Syntesizes its particles

2. Laboratory diagnosis of viruses Laboratory diagnosis of viruses Indirect dg. Direct Microscopy light (only big viruses –POXVIRUS) electron whole virus ( Rotavirus ) inclusions ( Negri bodies – rabies ) PCR Immunofluorescence – with cultivation (shell vial, task 4) Isolation – animals, chicken embryo (10 days old), tissue culture ELISA (Rotavirus)

3. Animals in Virology  Mouse (HSV)  Suckling mouse babies (Coxsackie viruses, tick-born encefalitis)  Rabbit (cornea- herpes virus)

4. Chicken embryo (task 2) Yolc sac Y P..paper membrane Y..Yolc sac w..White A..air A Viruses used for inoculation of: Amnion…influenza Alantois…influenza for vaccine preparation CAM…Poxviruses, HSV Yolc sac…encephalitic viruses, Chlamydia, Rickettsia w Yolc sac CAM…chorioalantoid membrane Inoculation of : Amnion fluid

5. Tissue cultures (task 3) Used only 1xUsefull 2x – 10x Primary culturesDiploid c. Monkey kidneyHuman embryon. lungs (LEP) Indestructible - from tumours Cell culture (HELA cells)

6. Preparation of tissue cultures MEM (medium) + ATB+ cells from tissue Cultivation in Roux bottle + Sample Changes of cells (also medium colour) CPE (cytopatic efect) Other – not visible in microscope I. Plaques (hollows) II. Rounded cells III. Syncitia Monkey cells fibroblasts Determination of a virus Visualization Interference Hemadsorbtion adhesion of erytrocytes 1.Virus with CPE 2. Our virus Cells are growing without CPE

7. Indirect virus diagnosis (Ig) CFT (ag+ab + C + Indicator system)CFT (ag+ab + C + Indicator system) no haemolysis haemolysis (2 samples are needed) – task 5 ELISA Immunofluorescence Western blott HITHIT (a.b. adhere on virus and stop agglutination) – possible only in hemagglutination viruses – task 6 Neutralisation + -

8. Shell vial (task 4)-rapid cultivation – in life threating TC on small glass in glass test-tube + sample (CSF) centrifugation 1 d cultivation t 37 °C Remove of a cover glass, stick to under glass Indirect IF (CMV) Direct IF (Herpes 1,2) under glass cover glass + Ag a.b. marked with an fluorescence colour - conjugate Monoclonal a.b. Conjugate shining Sample is shining