microbiology labwork jpems 2012 nantes dr c. bressollette (virology, nantes) dr j. caillon...
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MICROBIOLOGY LABWORKJPEMS 2012
NANTES
Dr C. Bressollette (Virology, Nantes)Dr J. Caillon (Bacteriology, Nantes)
Dr M. Eveillard (Bacteriology, Angers)Cécile Peltier (Virology, Nantes)
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Safety rules
Wash your hands at the beginning and the end of the session Wear a labcoat Wear disposables gloves when recommended (IHA, PCR) For some of the bacteriology procedures, you will work around
the flame of a Bunsen burner > long hair must be restrained Throw the « infected material » in the biohazard container Clean small spills with bleach Disinfect work areas after use
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Virology– Viral cultures:
• Observation of a cytopathic effect• HSV typing by immunofluorescence
– Molecular assay• Amplification of a viral gene
– Serologic assay: • Inhibition of hemagglutination
Bacteriology Analysis of a bacterial mixture
Gram staining
Bacterial identification
Susceptibility to antibiotics
Analysis of the environment
The week schedule
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Day 1 - Virology
The laminar flow hood / biosafety hood
Plastic culture flasks
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Day 1 - Virology
Observation of non infected adherent cells under the microscope
Human fibroblasts Primary cells, limited multiplication in vitro
HEP-2: epithelial cells, immortalized cell line
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Day 1 - Bacteriology
Study of a mix of two bacterial strains 6 different bacterial mixtures6 different bacterial mixtures:: A, B, C, D, E : mark the letter A, B, C, D, E : mark the letter
on your mix tubeon your mix tube Microscopic observation
– Smear colored by the method of gram staining
• See the demonstration and technique
• Pink bacteria: gram negative
• Blue bacteria: gram positive
Bacterial isolation on an agar medium (quadrant method)
• See the demonstration and your workbook
• 3 plates– Chromogenic media– Drigalski plate (green)– Chapman plate (pink):