TABLE I
Routine Tests for Adventitious Viruses
| GENERAL VIRUS DETECTION ASSAYS |
|---|
 In vitro cell culture tests |
| • Inoculation of three monolayer culture types |
| – same species |
| – human diploid cells |
| – monkey kidney cells |
| • Detect cytopathic viruses based upon cytopathic effect (CPE) |
| • Detect non-cytopathic viruses by testing for hemadsorption or hemagglutination of RBCs |
| – human (type O), mammalian, and avian species |
| – at least at two different temperatures |
 In vivo assays |
| • Inoculation of different animals species using different routes |
| – Adult mice |
| – Suckling mice |
| – Embryonated hens' eggs |
| – Guinea pigs, rabbits (in some cases) |
| • For mammalian species, animals are observed for change in health status. Cause of death is determined by gross pathology, histopathology and if virus infection is suspected, identification is attempted. |
| • For hens' eggs, tests are done for hemagglutinating agents with RBCs from guinea pigs, humans (type O), and an avian species. |
 Transmission electron microscopy (TEM) |
 Reverse transcriptase assay for retroviruses (PERT) |
| SPECIES-SPECIFIC VIRUS ASSAYS |
 Tests for animal viruses (based upon source materials) |
| • Culture assay on appropriate indicator cells followed by virus-specific fluorescent detection assays per 9CFR 113.47 and 113.53 |
 Antibody production assays for rodent viruses (MAP, HAP, RAP) |
| • Injection of test article into SPF mouse, hamster, rat |
| • Assay for presence of antibody to specific viruses by ELISA, hemagglutination inhibition, and IFA |
| • MAP should include LCMV challenge. |
 PCR or infectivity assays for known viruses |
| • Rodent cells: retroviruses, LCMV |
| • Simian cells: retroviruses, polyomaviruses |
| • Human cells: retroviruses, hepatitis viruses, enteroviruses, papillomaviruses, polyomaviruses, adenoviruses, herpesviruses |