Mouse antibody production test: can we do without it?

Area of Science:

• Veterinary Virology
• Laboratory Animal Science
• Diagnostic Microbiology

Background:

• Microbiological contamination in experimental mice can lead to infections and compromise research integrity.
• Sensitive, reliable, and routine diagnostic methods are essential for screening biological materials used in animal research.

Purpose of the Study:

• To compare the sensitivity of three common diagnostic assays: viral plaque assay, mouse antibody production (MAP) test, and polymerase chain reaction (PCR).
• To evaluate these methods for the detection of two prevalent murine viruses: Mouse Hepatitis Virus strain A59 (MHV-A59) and Murine Minute Virus (MMVp).

Main Methods:

• Comparative analysis of viral plaque assay, MAP test (day 28), and reverse transcription PCR (RT-PCR) for MHV-A59 detection.
• Comparative analysis of viral plaque assay, MAP test (day 28), and PCR for MMVp detection.
• Serial tenfold dilutions of virus stocks were used to determine assay sensitivity.

Main Results:

• For MHV-A59, the MAP test (10^-10 dilution) was 10-fold more sensitive than the plaque assay (10^-9) and 10^4-fold more sensitive than RT-PCR (10^-6).
• For MMVp, PCR (10^-10 dilution) was 10^6-fold more sensitive than both the plaque assay and MAP test (10^-4 dilution).
• Assay sensitivity varied significantly depending on the specific virus being targeted.

Conclusions:

• The choice of diagnostic method for murine viruses should be based on determined sensitivity for the specific virus of interest.
• The most sensitive detection method must be independently established for each virus to ensure accurate screening of biological materials.
• Implementing sensitive diagnostic protocols is vital for maintaining the health of experimental mouse colonies and the validity of research outcomes.