In vivo adventitious virus detection
Another method used to detect viruses is the inoculation of virus-sensitive animals with the test product via multiple routes. Four species are generally used, as they make it possible to screen for a wide range of viruses:
Embryonated eggs |
Suckling mice |
Adult mice |
Guinea pigs |
Arboviruses |
Arboviruses |
Arboviruses |
Arboviruses |
Herpes simplex type 2 |
Coxsackie A Virus |
Herpes simplex type 1 |
Ebola |
Influenza |
Coxsackie B Virus |
Herpes simplex type 2 |
Encephalomyocarditis |
Mumps |
Foot and mouth disease virus |
Lassa |
Junin |
Parainfluenza 1 |
Herpes simplex type 1 |
Rabies
|
Marburg |
Rabies |
Machupo |
|
|
Vaccinia |
Variola |
|
|
|
Rabies |
|
|
|
Vaccinia |
|
|
Texcell can provide you with four separate standard protocols or compiled protocols involving different animals, depending on your needs.
Protocol 0831: Detection of viruses by using embryonated chicken eggs |
Product to be tested: cell banks, unprocessed bulks |
This method is used to determine whether a viral contaminant is present in the test article by inoculation of chicken eggs, followed by examination of the embryo survival. At least 10 eggs for each route (intra-amniotic, intra-allantoic and intraveittelic) are inoculated with the test article. For eggs from intra-allantoic and intra-vittellic routes, a passage is carried out at day+3 and day+9 respectively. For the intra-allantoic and intra-amniotic routes, haemagglutination tests are performed with egg fluids at the end of each incubation period. |
|
Protocol 0832: Detection of viruses by using suckling mice |
Product to be tested: cell banks, unprocessed bulks |
This method is used to determine whether a viral contaminant is present in the test article by inoculation of two groups of 10 suckling mice via three different routes (intramuscularly, intraperitoneally and intracerebrally), followed by observation of the animals (two 14-days periods). Mouse survival is checked at the end of the test. Dead mice are autopsied in order to observe any pathological signs. |
|
Protocol 0833: Detection of viruses by using adult mice |
Product to be tested: cell banks, unprocessed bulks |
This method is used to determine whether a viral contaminant is present in the test article by inoculation of two groups of 10 adult mice via three different routes (intramuscularly, intraperitoneally and intracerebrally), followed by observation of the animals over 14 days. Mouse survival is checked at the end of the test. Dead mice are autopsied in order to observe any pathological signs. |
|
Protocol 0834: Detection of viruses by using guinea pigs |
Product to be tested: cell banks, unprocessed bulks |
This method is used to determine whether a viral contaminant is present in the test article by inoculation of two groups of guinea pigs respectively 5 for the test article and 4 for negative controls, via two different routes (intraperitoneally and intracerebrally), followed by observation of the animals over 28 days. Guinea pig survival is checked at the end of the test. Dead guinea pigs are autopsied in order to observe any pathological signs. |
|
Protocol N° 0836 (a compilation of protocols N° 0831, 0832, 0833 and 0834) is also available.
Protocol N° 0837 (a compilation of protocols N° 0831, 0832 and 0833) is also available.
For the evaluation of attenuated virus vaccines, the European Pharmacopoeia (EP) assays are generally used:
Protocol 0843: Detection of viruses by using suckling mice (EP) |
Product to be tested: cell banks, unprocessed bulks |
This method is used to determine whether a viral contaminant is present in the test article by inoculation of two groups of 10 suckling mice (10 controls and 10 samples) via two different routes, (intraperitoneally (0.1 ml) and intracerebrally (0.01 ml)), followed by observation of the animals over 14 days. If animals die or show signs of illness, a subinoculation on 5 mice is carried out and observation over 14 days is performed. The validity of the test is confirmed if at least 80% of the mice survive. |
|
Protocol 0844: Detection of viruses by using adult mice (EP) |
Product to be tested: cell banks, unprocessed bulks |
This method is used to determine whether a viral contaminant is present in the test article by inoculation of two groups of 10 adult mice (10 controls and 10 samples) via two different routes (intraperitoneally (0.5 ml) and intracerebrally (0.03 ml)), followed by observation of the animals over 14 days. If animals die or show signs of illness, a subinoculation on 5 mice is carried out and an observation over 14 days is performed. The validity of the test is confirmed if at least 80% of the mice survive. |
|
Protocol 0845: Detection of viruses by using guinea pigs (EP) |
Product to be tested: cell banks, unprocessed bulks |
This method is used to determine whether a viral contaminant is present in the test article by inoculation of two groups of guinea pigs (4 controls and 5 samples) via the intraperitoneal route (5 ml), followed by observation of the animal over 42 days. Guinea pig survival is checked at the end of the test. The validity of the test is confirmed if at least 80% of the guinea pigs survive. |
|
Protocol N° 0847 (a compilation of protocols N° 0843, 0844 and 0845) is also available.
