Objective      To establish a method for detection of murine hepatitis virus (MHV) by reverse dot blotting (RDB).       Methods      L929 cells were infected with 4 MHV strains separately. MHV was harvested and RNA was extracted for reverse transcription to obtain cDNA. Primers and specific probes were designed  on the basis of conserved sequence of MHV and forward primer was labeled with biotin. The method of RDB was developed using polymerase chain reaction (PCR) products. The stability

specificity and sensitivity of RDB were evaluated. In addition

41 mice we

re tested by RDB and ELISA. The positive samples tested by RDB were detected by cloning and sequencing.       Results      The test strip of RDB could be kept at 4 ℃ for 8 months. The specificity of RDB was 100% for MHV (when testing with

Salmonella sp

.

Staphylococcus aureus

and hepatitis B virus) and the limit of detection was down to 5 ng/μL MHV PCR products. In 41 mice tested by RDB

there were 5 MHV-positive mice distributed in three groups. But by ELISA

there were five MHV-positive mice distributed in 2 groups. The results by cloning and sequencing were consistent with the positive samples tested by RDB.       Conclusions      RDB is simple

reliable

sensitive and specific for identification of MHV of laboratory animals.