Virus stocks
Virus assays
Decontamination and cleaning systems
Storage systems
Virus stocks
Using qualified high-titre virus stocks; typically 7-8 log TCID50/ml
Virus available as serum-containing and serum-free stocks; typically non-concentrated virus culture supernatant is used
Undetectable aggregation of CPV, PPV, and HAV stocks as revealed by 35 nm filtration studies (Planova 35N)
Undetectable aggregation of BVDV, HIV, and PSR stocks as revealed by 0.22 µm filtration studies
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Virus assays
Samples are tested within 10 minutes after collection
Minimal handling of samples after collecting; e.g. cytotoxicity of samples is stopped by an one-step dilution in culture medium; so no ultra-centrifugation or column steps, and no freezing and thawing of samples prior to testing
TCID50 assays testing 3-fold serial dilutions in 8 or more replicates
Bulk culture tests used for ultra sensitive detection of viruses
Extended culture times to prevent false negative results
| Virus | Repeatability | Reproducibility |
Mean titre
[log TCID50/ml] | 95% CL | Mean titre
[log TCID50/ml] | 95% CL |
| BVDV | 6.38 | 0.15 | 6.42 | 0.18 |
| CPV | 7.28 | 0.15 | 7.32 | 0.22 |
| EMC | 7.60 | 0.13 | 7.55 | 0.15 |
| HAV | 6.93 | 0.10 | 7.13 | 0.34 |
| HIV | 7.93 | 0.23 | 7.93 | 0.20 |
| PPV | 7.67 | 0.13 | 7.74 | 0.22 |
| PSR | 8.48 | 0.15 | 8.73 | 0.21 |
| SV40 | 7.28 | 0.09 | 7.19 | 0.19 |
| VSV | 6.31 | 0.08 | 6.36 | 0.10 |
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Decontamination and cleaning systems
Equipment used is decontaminated with a validated sanitization procedure and (if possible) autoclaved when exported from the laboratory
Solid waste is contained and decontaminated by autoclaving with a biohazard destruction program
Liquid waste is collected in a container and decontaminated with a validated heat treatment procedure
The facility is cleaned weekly by laboratory personnel and subsequently by a certified cleaning service
Twice a year, the complete facility is extensively cleaned by the cleaning service
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Storage systems
Qualified master, production, and working virus stocks are stored in -80°C freezers
In case of a technical failure of a -80°C freezer, a liquid nitrogen back-up system is available to assure the temperature of the stored materials
A part of master virus stocks is stored in a -80°C freezer located in a separate building
Qualified master and production cell banks are stored in a biostate located outside the facility at the Sanquin Department of Cryobiology
As a back up part of the master cell banks is stored in a biostate located in a building outside Sanquin
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