IMPACT: PCR-based Alternative to MAP testing
The Infectious Microbe PCR Amplification Test, or IMPACT, is a panel of PCR assays that detects murine pathogens in biological samples as an alternative to the Mouse Antibody Production (MAP) test. Contamination of biological specimens, such as cell lines, hybridomas and tumor cells, with rodent pathogens can result in devastating outbreaks of disease in laboratory animals implanted with these materials and confounding and deleterious effects on tissue culture-based experiments. The traditional method for testing biological specimens for murine pathogens has been the Mouse Antibody Production (MAP) test. The major disadvantage of MAP testing is the 6 to 8 weeks required to get results. Comparison of IMPACT results with MAP testing results for representative DNA and RNA viruses indicated that the sensitivity of the IMPACT was equal or greater than that of MAP testing. Turnaround time for IMPACT results is usually 10 business days.
Sample Collection and Preparation for Shipping
IMPACT: To test biological specimens or cultured cells by the IMPACT, send two 2ml cryovials of each sample with 1 x 107 cells/vial in 1 ml of growth media or phosphate-buffered saline to our laboratory on a minimum of 2 kg of dry ice by overnight courier.
Ascites fluids: Ascites fluids for PCR evaluation may be prepared in one of two ways. Freshly collected ascites fluid should be shipped cooled (not frozen) with wet ice in a styrofoam container by overnight service. Because of the limited number of cells that may be present in ascites, we recommend that 10 ml of fluid be submitted. Alternatively, cells in the fluids can be pelleted by centrifugation at 10,000 x g for 10 minutes, the supernatant removed, and the cell pellet frozen. The pelleted sample should be shipped on dry ice in a styrofoam container by an overnight carrier that handles dry ice shipments.
Tissue or cellular samples: Collection of tissues or cells for PCR evaluation should be performed aseptically to prevent inadvertent contamination of the samples. Each cellular sample should be placed in individually labeled sterile containers. Instruments used for collection should be replaced between samples. Alternatively, instruments can be wiped clean and immersed in a diluted bleach (10 percent) solution for 10 minutes prior to reuse. Tissue samples should be frozen and shipped on dry ice in a styrofoam container by an overnight carrier service that handles dry ice shipments.
|Mouse IMPACT (16)||Rat IMPACT (13)|
|Mycoplasma spp.||Mycoplasma spp.|
|Sendai virus||Sendai virus|
|Pneumonia virus of mice||Pneumonia virus of mice|
|Lymphocytic choriomeningitis virus||Lymphocytic choriomeningitis virus|
|Mouse adenovirus (MAD1, MAD2)||Mouse adenovirus (MAD1, MAD2)|
|Mouse parvovirus||Rat parvovirus|
|Minute virus of mice||Minute virus of rats|
|Mouse hepatitis virus||Kilham's rat virus|
|Theiler's murine encephalomyelitis virus||Toolan's H1|
|Mouse norovirus||Rat Coranavirus|
|Lactate dehydrogenase-elevating virus|