Testing of Viruses Flashcards
Conventional cell culture cell monolayer
Growing cell monolayer (single confluent layer) on plastic or glass tube
Conventional cell culture incubation conditions
Roller drum, tilted 5-7 degrees, revolving at 0.5-1 rpm at 35-37°C for 1 to 4 weeks
Advantage of conventional cell culture
Can detect different viruses in one cell culture
Disadvantage of conventional cell culture
Long, laborious process
EDTA/Heparin blood handling
Separate buffy coat
CSF specimen handling
Inoculate 1 mL directly
Feces/Rectal swab handling
Place in 2 mL VTM → Vortex → Centrifuge → Inoculate
NPS/OPS/Sputum/Respiratory washings handling
Dilute with VTM
Tissue specimen handling
Mince with scalpel/scissors → Gently grind → Suspend in VTM → Centrifuge → Inoculate
Swab/Fluid specimen handling
Emulsify in VTM
Urine specimen handling (clear)
Inoculate directly
Urine specimen handling (turbid)
Centrifuge → Inoculate
Genital/Skin (Vesicle fluid/Scrapings) specimen handling
Emulsify in VTM
Urine cell culture for adenovirus
HDF, HEp-2
Major cell culture types
PMK, HDF, HEp-2
Genital/Skin cell culture
HDF
Cell culture tube procedure
Absorption (12-24 hours) → Maintenance tube (replace 1-2x/week) → Incubation (5-28 days)
Freezing in cell culture
Remove old medium → Add fresh culture medium