Serological Analysis Of Blood Samples From Sentinel Rats

The pathogen status of the rats is most easily monitored by serological analysis of serum samples from sentinel rats, screening for the presence of serum antibodies to a panel of viral and bacterial microorganisms. Additionally, the animals should be routinely screened for common parasites (unit 1.1).

Materials

PBS (appendix 2)

Additional reagents and equipment for anesthesia by methoxyflurane inhalation (unit 1.4), tail vein or orbital plexus blood collection (unit 1.7), and preparation of serum from blood (unit 2.4)

1. Anesthetize each sentinel rat using methoxyflurane inhalation (unit 1.4).

2. Collect ~1 ml blood from the tail vein or orbital plexus (unit 1.7) of anesthetized sentinel rat into a 1.5-ml microcentrifuge tube.

4. Microcentrifuge tubes 10 min at 4000 rpm to pellet cells and collect serum for analysis.

5. Dilute 200 pl serum 1:5 in PBS and submit to Charles River Laboratories (see appendix 5) or one of the government-sponsored laboratories listed in Table 1.1.2 for complete serological assessment for rats.

A complete assessment includes screening for Sendai virus, pneumonia virus of mice (PVM), sialodacryoadenitis virus (SDAV), rat coronavirus, Kilham rat virus (KRV), H-1 virus, GD7, Reovirus-3, Mycoplasma pulmonis, lymphocytic choriomeningitis virus (LCMV; see unit 19.9), mouse adenovirus, Hantaan virus, Encephalitozoon cuniculi, and carbacillus. Monthly serological assessment of sentinel animals is recommended.

A new rat parvovirus with antigenic similarity to KRV) has recently been identified and designated RPV-1 (Jacoby and Ball-Goodrich, 1995). The RPV-1 status of the colony should also be monitored. At this writing, this can be done by requesting an additional immunofluorescence analysis for KRV; the presence of RPV-1 is indicated by a negative ELISA test for KRV and a positive immunofluorescence test for KRV.

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