Rabid
raccoon dogs (
Nyctereutes procyonoides koreensis) have been responsible for
animal rabies in
South Korea since the 1990s. A recombinant
rabies vaccine strain, designated as ERAGS, was constructed for use as a bait
vaccine. Therefore, new means of differentiating ERAGS from other
rabies virus (RABV)
strains will be required in
biological manufacturing and
diagnostic service centers. In this study, we designed two specific primer sets for differentiation between ERAGS and other RABVs based on
mutation in the RABV
glycoprotein gene.
Polymerase chain reaction analysis of the
glycoprotein gene revealed two
DNA bands of 383 bp and 583 bp in the ERAGS
strain but a single
DNA band of 383 bp in the field
strains. The
detection limits of multiplex
reverse transcription polymerase chain reaction (RT-PCR) were 80 and 8 FAID 50 /reaction for the ERAGS and Evelyn-Rokitnicki-Abelseth
strains, respectively. No crossreactions were detected in the non-RABV reference
viruses, including
canine distemper virus,
parvovirus, canine adenovirus type 1 and 2, and
parainfluenza virus. The results of multiplex RT-PCR were 100% consistent with those of the fluorescent antibody test. Therefore, one-step multiplex RT-PCR is likely useful for differentiation between RABVs with and those without
mutation at position 333 of the RABV
glycoprotein gene.