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Experiment. 1. Hemagglutination test 2. Hemagglutination inhibition test 3. Observation morphology of fungi. Ⅰ. Hemagglutination test. Principle: Some HA-bearing viruses agglutinate RBCs in vitro. discard 0.25ml Tubes 1 2 3 4 5 6 7
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Experiment 1. Hemagglutination test 2. Hemagglutination inhibition test 3. Observation morphology of fungi
Ⅰ. Hemagglutination test • Principle:Some HA-bearing viruses agglutinate RBCs in vitro.
discard 0.25ml Tubes1 2 3 4 5 6 7 NS0.25 0.25 0.25 0.25 0.25 0.25 0.25 virus 0.25 0.25 0.25 0.25 0.25 0.25 (1:4) (1:8) (1:16) (1:32) (1:64) (1:128) (1:256) 0.5%RBC 0.25 0.25 0.25 0.25 0.25 0.25 0.25 (negative control) mix, RT, 45min~1hr,observation Ⅰ. Hemagglutination test • Procedures:
Ⅰ. Hemagglutination test • Record the results
Ⅰ. Hemagglutination test • Interpretation • Hemagglutination titer: is determined as the highest dilution • ofvirusthat can cause an obvious ( ++) hemaglutination. 1 2 3 4 5 6 7 (1:8) (1:16) (1:32) (1:64) (1:128) (1:256) ++++ +++ ++ ++ + - control
Ⅱ. Hemagglutination inhibition test • Principle: • If a person is infected by HA-bearing virus, anti-HA Ab may appear in his serum. These antibodies can block hemagglutination and lead to the hemagglutination inhibition phenomenon.
Ⅱ. Hemagglutination inhibition test • Procedures: Discard 0.25ml Tube 1 2 3 4 5 6 7 8 NS(ml) 0.45 0.25 0.25 0.25 0.25 0.25 0.45 0.25 serum 0.05 0.25 0.25 0.25 0.25 0.25 0.05 (1:10) (1:20) (1:40) (1:80) (1:160) (1:320) Virus(4U) 0.25 0.25 0.25 0.25 0.25 0.25 0.25 0.5% RBC 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Mix, RT, 1hr, observation
Ⅱ. Hemagglutination inhibition test • Interpretation: • Hemagglutination inhibition titer is determined as the highest dilution of serum that can completely inhibit hemagglutination.
Ⅱ. Hemagglutination inhibition test 1 2 3 4 5 6 7 8 Acute phase 1 2 3 4 5 6 7 8 Convalescent phase 1:10 1:20 1:40 1:80 1:160 1:320 - +
Ⅲ. Observation morphology of fungi • Observe colony morphology of three species of fungi • Observe morphology of fungi under a light microscope
Ⅲ. Observation morphology of fungi A. Observe colony morphology of three species of fungi 1.Sabouraud's agar plate culture of Cryptococcus neoformans ---yeast colony 2. Sabouraud's agar plate culture of Candida albicans --- yeast-like colony 3. Sabouraud's agar plate culture of mold --- filamentous colony
Yeast colony Ⅲ. Observation morphology of fungi
Ⅲ. Observation morphology of fungi Filamentous colony
Ⅲ. Observation morphology of fungi B. Observe morphology of fungi under a light microscope 1. India ink smear of Cryptococcus neoformans: capsules 2. Microcultures of Candida albicans: pseudohypha and chlamydospores 3. Microculture of mold: hypha and spores
Ⅲ. Observation morphology of fungi Cryptococcus neoformans
Ⅲ. Observation morphology of fungi Candida albicans