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Laboratory Diagnosis of Viruses and Cultivation Methods

This article provides an overview of laboratory diagnosis methods for viruses, including microscopy, PCR, immunofluorescence, and ELISA, as well as different techniques for virus cultivation such as tissue culture and inoculation in animal models. It also discusses the visualization of virus effects and indirect virus diagnosis methods.

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Laboratory Diagnosis of Viruses and Cultivation Methods

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  1. Viruses • Only1 NA • Small • Syntesizesitsparticles

  2. Laboratorydiagnosis of viruses Indirect dg. Direct • Microscopylight (onlybig viruses –POXVIRUS) electron whole virus (Rotavirus) inclusions (Negri bodies – rabies) • PCR • Immunofluorescence – withcultivation (shell vial, task 4) • Isolation– animals, chickenembryo (10 days old), tissueculture • ELISA(Rotavirus)

  3. AnimalsinVirology • Mouse (HSV) • Suckling mouse babies (Coxsackie viruses, tick-born encefalitis) • Rabbit (cornea- herpes virus)

  4. Chickenembryo (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

  5. Tissuecultures (task 3) Usedonly 1xUsefull 2x– 10x Primarycultures Diploid c. MonkeykidneyHuman embryon. lungs (LEP) Indestructible- fromtumours Cellculture (HELA cells)

  6. Preparation of tissue cultures Changesof cells (alsomedium colour) MEM (medium) + ATB+ cellsfrom tissue + Sample Other – not visible in microscope CPE (cytopatic efect) • I. Plaques (hollows) • II. Rounded cells • III. Syncitia Visualization Cultivationin Roux bottle Hemadsorbtionadhesion of erytrocytes Interference 1.Virus with CPE 2. Our virus Monkeycells fibroblasts Determination of a virus Cellsare growingwithout CPE

  7. Indirectvirus diagnosis (Ig) + - • CFT (ag+ab + C + Indicator system) no haemolysishaemolysis (2 samples are needed) – task 5 • ELISA • Immunofluorescence • Western blott • HIT (a.b. adhereon virus andstop agglutination) – possible only in hemagglutination viruses – task 6 • Neutralisation

  8. Shell vial (task 4)-rapidcultivation – in life threating centrifugation Remove of acoverglass, stick tounderglass + sample (CSF) 1 d cultivationt 37 °C TCon small glassin glass test-tube Indirect IF (CMV) Direct IF (Herpes 1,2) Sampleis shining Conjugate a.b. marked withan fluorescence colour-conjugate Monoclonala.b. coverglass + Ag underglass

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