Viruses Only 1 NA Small Syntesizes its particles
Laboratory diagnosis of viruses Indirect dg. 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)
Animals in Virology Mouse (HSV) Suckling mouse babies (Coxsackie viruses, tick-born encefalitis) Rabbit (cornea- herpes virus)
Chicken embryo (task 2) Inoculation of : Amnion fluid Yolc sac CAM…chorioalantoid membrane A Viruses used for inoculation of: Amnion…influenza Alantois…influenza for vaccine preparation CAM…Poxviruses, HSV Yolc sac…encephalitic viruses, Chlamydia, Rickettsia A..air Yolc sac Y P..paper membrane Y..Yolc sac w..White w
Tissue cultures (task 3) Used only 1x Usefull 2x – 10x Primary cultures Diploid c. Monkey kidney Human embryon. lungs (LEP) Indestructible - from tumours Cell culture (HELA cells)
Preparation of tissue cultures Changes of cells (also medium colour) MEM (medium) + ATB+ cells from tissue + Sample Other – not visible in microscope CPE (cytopatic efect) I. Plaques (hollows) II. Rounded cells III. Syncitia Visualization Cultivation in Roux bottle Hemadsorbtion adhesion of erytrocytes Interference 1.Virus with CPE 2. Our virus Monkey cells fibroblasts Determination of a virus Cells are growing without CPE
Indirect virus diagnosis (Ig) CFT (ag+ab + C + Indicator system) no haemolysis haemolysis (2 samples are needed) – task 5 ELISA Immunofluorescence Western blott HIT (a.b. adhere on virus and stop agglutination) – possible only in hemagglutination viruses – task 6 Neutralisation + -
Shell vial (task 4)-rapid cultivation – in life threating centrifugation Remove of a cover glass, stick to under glass + sample (CSF) 1 d cultivation t 37 °C TC on small glass in glass test-tube Indirect IF (CMV) Direct IF (Herpes 1,2) Sample is shining Conjugate a.b. marked with an fluorescence colour - conjugate Monoclonal a.b. cover glass + Ag under glass