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Results in 5 Business Days |
Individual PCR Tests
Mycoplasma Testing
PCR Parvovirus Panels
PCR-based Alternative to MAP testing
IMPACT Profiles
h-IMPACT Profile
Information on Sample Collection
MU RADIL offers a variety of PCR assays to aid in detection of infectious agents in tissues, cell lines and other types of biologic specimens.
PCR assays are available to detect bacterial, viral and protozoal pathogens in animal tissues. In addition, our laboratory developed the Infectious Microbe PCR AmplifiCation Test or IMPACT, which is a panel of PCR assays that detects murine pathogens in biological samples as an alternative to the Mouse Antibody Production (MAP) test. Numerous types of biologic materials can be evaluated by PCR assays.
h-IMPACT - Testing of biological specimens for human pathogens.
Contamination of biological specimens of human origin, such as human cell lines
and tumors, with human pathogens can pose a health risk to personnel handling the specimens.
Contaminated specimens can also add unwanted research variables. RADIL has developed and validated a panel of
PCR assays, called h-IMPACT, for testing biological specimens for the presence of the
following human pathogens: Human immunodeficiency virus 1 (HIV1), Human Immunodeficiency virus 2 (HIV2),
Hepatitis A virus, Hepatitis B virus, Hepatitis C virus, Sin Nombre virus, Hantaan virus and Seoul virus. In addition, the panel
will detect Mycoplasma spp., common contaminants of cell cultures. Results to be used for research purposes only.
If you have questions about the most appropriate sample to submit or how to collect the specimen, call or e-mail the RADIL. Our PCR Services are expanding rapidly; we recommend that you call or e-mail the laboratory if the test you desire is not included in the list of individual PCR tests.
With our on-line access to reports, results can be reviewed as soon as testing is completed and approved. E-mail notifications will be sent whenever results indicate a potential problem, as well as when the case is finalized.
Mycoplasma Testing
Mycoplasma contamination of cell lines can invalidate research investigations. In cell lines, virtually every cell process can be altered by Mycoplasma infection. Documented affects include:
- Diminished cell growth
- Altered cellular metabolism
- Phenotypic changes
- Increased cell-mediated cytotoxicity
- Altered mitogenicity of lymphocytes
- Interferon induction in tumor cell lines
- Diminished immunoglobulin production by B-cells
Detrimental effects on Embryonic Stem (ES) cells used for creation of genetically-modified mice have also been recognized. Mycoplasma contamination can:
- Reduce the number of chimeras generated
- Decrease pregnancy rates
- Reduce litter size
Mycoplasma contamination of cell lines and ES cells is a widespread problem. The prevalence of Mycoplasma contamination in cell lines and ES cells is high. Approximately 10% of all cell lines tested in our laboratory are positive for Mycoplasma sp. Mycoplasmas belong to the class Mollicutes, which includes over 100 species of Mycoplasma. The most prevalent Mycoplasma contaminants of cultured cells are Mycoplasma orale (human-origin), Mycoplasma hyorhinis (porcine-origin), Mycoplasma arginini (bovine-origin) and Acholeplasma laidlawii (bovine-origin).
Mycoplasma detection using a highly sensitive PCR assay The RADIL offers a PCR assay that is exquisitely sensitive with the capability of detecting a single genomic copy of Mycoplasma sp. The assay detects all species of Mycoplasma known to infect cell cultures. The assay also detects Acholeplasma laidlawii and Spiroplasma mirum, two other members of the Mollicutes class that also can contaminate cultured cells.
To submit cultured cells for Mycoplasma testing Send one cryovial of each sample containing ~ 2x106 – 1 x 107 cells. Cells for testing may be submitted in several formats. Cells may be pellets of cells in growth medium or phosphate buffered saline. Alternatively, cells may be vials of frozen cells with cryoprotectant. Please ship samples to our laboratory frozen via overnight courier with sufficient ice packs in the container that samples remain frozen during shipment as thawing can be detrimental to the accuracy of the testing results.
Individual PCR Tests
Biological reagents (e.g. fluids, cultured cells, tumors) and samples (such as organ biopsies, feces, swabs) can be tested by PCR for the following list of micobial agents. Please note: the specimens listed in parentheses are optimal for submission for PCR testing.
| VIRUSES | BACTERIA | PROTOZOA | OTHER |
|---|---|---|---|
| Adenovirus: MAD1 (lung), MAD2 (intestine), GpAD (lung) | Cilia-associated respiratory bacillus (trachea, lung) | Encephalitozoon cuniculi (urine, kidney, brain) | Pneumocystis carinii (lung) |
| Coronavirus: MHV (feces, mesenteric lymph node), RCV, SDAV (salivary, Hardarian gland) | Clostridium piliforme (cecum) | ||
| Cytomegalovirus: mCMV, rCMV, gpCMV (spleen, salivary gland) | Citrobacter rodentium (feces, cecum) | ||
| Ectromelia (spleen, skin lesions) | Corynebacterium bovis (skin scrapings, skin) | ||
| Hantavirus: Hantaan, Seoul, Puumala (kidney), Sin Nombre (lung) |
Helicobacter (feces, cecum) includes H. bilis, H. hepaticus, H. rodentium, H. trogontum, H. typhlonius, H. species |
||
| K virus (kidney) | Mycoplasma pulmonis (nasopharynx, trachea) | ||
| Lactate dehydrogenase-elevating virus (spleen) | Mycoplasma spp. (all known species) (cell cultures) | ||
| Lymphocytic choriomeningitis virus (kidney) | Pasteurella sp.(oral cavity, uterus) | ||
| Mouse thymic virus (salivary gland, thymus) | Proteus sp. (feces, cecum) | ||
| Murine norovirus, MNV (mesenteric lymph node, spleen, feces) | Pseudomonas aeruginosa (feces, cecum) | ||
| Parvovirus: Generic, MVM, MPV, KRV, H-1, RPV, RMV (mesenteric lymph node, spleen) | Salmonella sp. (feces, cecum) | ||
| Pneumonia virus of mice (trachea, lung) | Staphylococcus aureus (nasopharynx) | ||
| Polyoma virus (skin, mammary gland) | |||
| Reovirus 3 (liver, lung, feces) | |||
| Rotavirus (EDIM) (feces, intestine) | |||
| Sendai virus (trachea, lung) | |||
Theiler's encephalomyelitis virus (feces, intestine) |
PCR Parvovirus Panels
| Mouse $77.25* |
Rat $77.25* |
|---|---|
| MVM (MPV1, MPV2, MPV3) | KRV, RMV, H-1, RPV |
* A 5% discount is applied to each case with 5 or more samples.
A 10% discount is applied to each case wtih 10 or more samples.
IMPACT: PCR-based Alternative to MAP testing
RADIL developed and validated a PCR-based alternative to MAP testing. 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. To address this issue, our laboratory has developed the Infectious Microbe PCR AmplifiCation Test or IMPACT , which is a panel of PCR assays that detects murine pathogens. 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 in 5 business days. The cost of testing by the IMPACT is markedly lower than commercial MAP testing.
IMPACT Profiles - Mice
|
IMPACT Profile I
(22 Agent Tests) $456.50* |
IMPACT Profile II
(19 Agent Tests) $394.40* |
IMPACT Profile III
(16 Agent Tests) $358.25* |
IMPACT Profile IV
(9 Agent Tests) $185.50* |
|---|---|---|---|
| Mycoplasma spp. | Mycoplasma spp. | Mycoplasma spp. | Mycoplasma spp. |
| Sendai virus | Sendai virus | Sendai virus | Sendai virus |
| Mouse hepatitis virus | Mouse hepatitis virus | Mouse hepatitis virus | Mouse hepatitis virus |
| Pneumonia virus of mice | Pneumonia virus of mice | Pneumonia virus of mice | Pneumonia virus of mice |
| Minute virus of mice | Minute virus of mice | Minute virus of mice | Minute virus of mice |
| Mouse parvovirus (MPV1, MPV2, MPV3) | Mouse parvovirus (MPV1, MPV2, MPV3) | Mouse parvovirus (MPV1, MPV2, MPV3) | Mouse parvovirus (MPV1, MPV2, MPV3) |
| Theiler's murine encephalomyelitis virus | Theiler's murine encephalomyelitis virus | Theiler's murine encephalomyelitis virus | Theiler's murine encephalomyelitis virus |
| Murine norovirus | Murine norovirus | Murine norovirus | |
| Reovirus 3 | Reovirus 3 | Reovirus 3 | |
| Mouse rotavirus | Mouse rotavirus | Mouse rotavirus | |
| Ectromelia virus | Ectromelia virus | Ectromelia virus | |
| Lymphocytic choriomeningitis virus | Lymphocytic choriomeningitis virus | Lymphocytic choriomeningitis virus | |
| Polyoma virus | Polyoma virus | Polyoma virus | |
| Lactate dehydrogenase-elevating virus | Lactate dehydrogenase-elevating virus | Lactate dehydrogenase-elevating virus | |
| Mouse adenovirus (MAD1, MAD2) | Mouse adenovirus (MAD1, MAD2) | ||
| Mouse cytomegalovirus | Mouse cytomegalovirus | ||
| K virus | |||
| Mouse thymic virus | |||
| Hantaan virus |
* A 5% discount is applied to each case with 5 or more samples.
A 10% discount is applied to each case wtih 10 or more samples.
IMPACT Profiles - Rats
|
IMPACT Profile V
(15 Agent Tests) $381.00* |
IMPACT Profile VI
(11 Agent Tests) $298.70* |
|---|---|
| Mycoplasma spp. | Mycoplasma spp. |
| Pneumonia virus of mice | Pneumonia virus of mice |
| Kilham's rat virus | Kilham's rat virus |
| Toolan's H1 virus | Toolan's H1 virus |
| Rat parvovirus | Rat parvovirus |
| Lymphocytic choriomeningitis virus | Lymphocytic choriomeningitis virus |
| Rat cytomegalovirus | Rat cytomegalovirus |
| Sendai virus | Sendai virus |
| Rat coronavirus | Rat coronavirus |
| Sialodacryoadenitis virus | Sialodacryoadenitis virus |
| Rat minute virus | Rat minute virus |
| Seoul virus | |
| Mouse adenovirus | |
| Reovirus 3 | |
| Theiler's murine encephalomyelitis like virus |
* A 5% discount is applied to each case with 5 or more samples.
A 10% discount is applied to each case wtih 10 or more samples.
IMPACT Profile – Hamsters
|
IMPACT Profile VII
(11 Agent Tests) $298.70* |
|---|
| Mycoplasma spp. |
| Pneumonia virus of mice |
| Kilham's rat virus |
| Toolan's H1 virus |
| Hamster parvovirus |
| Lymphocytic chorlomeningtis virus |
| Minute virus of mice |
| Theiler's murine encephalomyelitis virus |
| Sendai virus |
| Reovirus 3 |
| Hantaan virus |
* A 5% discount is applied to each case with 5 or more samples.
A 10% discount is applied to each case wtih 10 or more samples.
IMPACT Profile – Comprehensive Murine Panel
|
IMPACT Profile VIII
(31 Agent Tests) $659.00* |
|---|
| Mycoplasma spp. |
| Sendai |
| Mouse hepatitis virus |
| Pneumonia virus of mice |
| Minute virus of mice |
| Mouse parvovirus (MPV1, MPV2, MPV3) |
| Murine Norovirus |
| Reovirus 3 |
| Mouse rotavirus |
| Ectromelia virus |
| Lymphocytic choriomeningitis virus |
| Polyoma virus |
| K virus |
| Mouse adenovirus (MAD1, MAD2) |
| Mouse cytomegalovirus |
| Lactate dehydrogenase-elevating virus |
| Mouse thymic virus |
| Hantaan virus |
| Kilham’s rat virus |
| Toolan’s H1 virus |
| Rat parvovirus |
| Rat cytomegalovirus |
| Rat coronavirus |
| Rat minute virus |
| Sialodacryoadenitis virus |
| Seoul virus |
| Theiler’s murine encephalomyelitis virus |
| Theiler’s murine encephalomyelitis like virus |
* A 5% discount is applied to each case with 5 or more samples.
A 10% discount is applied to each case wtih 10 or more samples.
h-IMPACT Profile
|
h-IMPACT Profile
(9 Agent Tests) $231.75* |
|---|
| Human immunodeficiency virus (HIV1, HIV2) |
| Hepatitis viruses (A, B and C) |
| Hantaviruses (Hantaan, Seoul, Sin Nombre) |
| Mycoplamsa spp. |
* A 5% discount is applied to each case with 5 or more samples.
A 10% discount is applied to each case wtih 10 or more samples.
Sample Collection and Preparation for Shipping
IMPACT To test biological specimens or cultured cells by the IMPACT, send two 2 ml cryovials of each sample with 1 x 107 cells/vial. Cells may be in the form of a pellet or in growth media, freeze media or phosphate-buffered saline. To test liquid samples by the IMPACT, send two 2 ml cryovials of each sample with 0.5 ml of sample/vial. Collection of material for IMPACT testing should be performed aseptically to prevent inadvertent contamination of the samples. Ship samples to our laboratory on a minimum of 2 kg of dry ice by overnight courier.
Mycoplasma: To test biological specimens or cultured cells for Mycoplasma sp. contamination, send one 1 ml cyrovial of each sample containing 2x106 to 1x107 cells/vial. Cells may be in the form of a pellet or in growth media, freeze media or phosphate-buffered saline. Collection of material for testing should be performed aseptically to prevent inadvertent contamination of the samples. Ship samples to our laboratory on a minimum of 2 kg of dry ice by overnight courier.
Feces: Fecal pellets for PCR evaluation should be collected with sterile forceps and placed in individually labeled sterile containers. Two to three pellets per animal are generally adequate. If multiple animals of the same microbiologic status are to be evaluated, up to 10 fecal pellets can be pooled and tested as one sample. If fecal pellets from multiple animals are to be collected, forceps should be replaced between animals to prevent cross-contamination. Alternatively, forceps can be wiped clean and immersed in a dilute bleach (10%) solution for 10 minutes prior to reuse. Fecal pellets for identification of viruses should be frozen and shipped on a minimum of 2 kg of dry ice in a styrofoam container by an overnight carrier service that handles dry ice shipments. Fecal pellets for identification of Helicobacters should be shipped by an overnight carrier, but do not need to be chilled or frozen.
Tissue samples: Collection of tissues for PCR evaluation should be performed aseptically to prevent inadvertent contamination of the samples. Each sample should be placed in individually labeled sterile containers. When collecting samples, instruments used to collect the samples should be replaced between samples. Alternatively, instruments can be wiped clean and immersed in a dilute bleach (10%) solution for 10 minutes prior to reuse. Tissue samples should be frozen and shipped on a minimum of 2 kg dry ice in a styrofoam container by an overnight courier.
See our Shipping Information for general shipping information. Call 800-669-0825 or e-mail the RADIL if you have questions about sample collection, storage, or submission.