MICROBIOLOGY LABWORKJPEMS 2012

NANTES

Dr C. Bressollette (Virology, Nantes)Dr J. Caillon (Bacteriology, Nantes)

Dr M. Eveillard (Bacteriology, Angers)Cécile Peltier (Virology, Nantes)

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

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

Day 1 - Virology

The laminar flow hood / biosafety hood

Plastic culture flasks

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

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):