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PROTOCOL FOR
RODENT ANTIBODY PRODUCTION (RAP) TESTS |
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| Rodent
Antibody Production, or RAP, tests are used routinely to
evaluate biological materials that are to be inoculated into rodents. This
is done to identify biological materials that may be contaminated with
infectious organisms that, when inoculated into a rodent, may cause an outbreak
of infectious disease within the animal facilities. This can result in the
elimination of entire rodent colonies, which can effectively shut down research
programs for significant periods of time. Two recent outbreaks emphasize
this fact: an outbreak of ectromelia in mice that was traced to inoculation of
contaminated serum, and a lymphocytic choriomeningitis virus (LCMV) outbreak
that was traced to inoculation of nude mice with contaminated tumor cell lines.1,2
Both outbreaks resulted in elimination of the infected mouse colonies, and the
LCMV outbreak resulted in infection of 10% of the personnel exposed to the
infected mice. The methods that are proposed here are consistent with current standards in laboratory animal medicine. Cell lines, transplantable tumors, hybridomas (used for monoclonal antibody production), embryonic stem cell lines, blood, blood products, and/or tissue homogenates may be tested if they were obtained from (or have been passed through) rodents of unknown disease status. This might include any human-origin tumor cell lines that have previously been passed through a rodent of unknown disease status. The procedure is as follows. Approximately 0.5 to 1.0 ml of crude lysate (prepared by several freeze-thaw cycles to lyse any cells that are present) is inoculated into each of two rodents of the same species (mice, rats, or hamsters) the investigator will be inoculating with the biological material. The rodents that are inoculated will be obtained from colonies that have repeatedly tested negative for rodent viruses and Mycoplasma pulmonis. The inoculum is administered by the intranasal, oral, and intraperitoneal routes and the rodents are housed under biocontainment conditions for approximately 4 weeks (blood is also collected ante-mortem from mice at 3 days post-inoculation to evaluate for the presence of LDEV by serum chemistry analysis). The rodents are then euthanized, blood and tissues are collected, and these samples are analyzed for evidence of viral and M. pulmonis infection by serology. The following agents have been documented as cell culture and/or transplantable tumor contaminants: lactase dehydrogenase elevating virus (LDEV), rodent coronaviruses (MHV and RCV/SDAV), rodent parvoviruses (MVM, H-1, KRV, MPV), ectromelia virus (mousepox), lymphocyctic choriomeningitis virus (LCMV), Theiler’s meningoencephalitis virus (TMEV), polyomavirus, mouse adenovirus, reovirus-3, and M. pulmonis. A recent study indicates that LDEV (a mouse-specific virus) is by far the most common contaminant detected in cell lines and transplantable tumors.3 Table 1 lists the infectious agents and fees for RAP testing for each rodent species. 1. Dick EJ Jr, Kittell CL, Meyer H, et al. (1996). Mousepox outbreak in a laboratory mouse colony. Lab. An. Sci. 46:602-611. 2. Dykewicz CA, Dato VM, Fisher-Hoch SP, et al. (1992). Lymphocytic choriomeningitis outbreak associated with nude mice in a research institute. J. Am. Med. Assoc. 267:1349-1353. 3. Nicklas W, Kraft V, and Meyer B. (1993). Contamination of transplantable tumors, cell lines, and monoclonal antibodies with rodent viruses. Lab. An. Sci. 43:296-300.
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