Clin Exp Vaccine Res.  2012 Jul;1(1):70-76. 10.7774/cevr.2012.1.1.70.

Development of a novel vaccine against canine parvovirus infection with a clinical isolate of the type 2b strain

• Park SA
• Park SY
• Song CS
• Choi IS
• Kim HY
• Lee JB
• Lee NH

• KMID: 2278790
• DOI: http://doi.org/10.7774/cevr.2012.1.1.70

Abstract

PURPOSE
In spite of an extensive vaccination program, parvoviral infections still pose a major threat to the health of dogs.
MATERIALS AND METHODS
We isolated a novel canine parvovirus (CPV) strain from a dog with enteritis. Nucleotide and amino acid sequence analysis of the isolate showed that it is a novel type 2b CPV with asparagine at the 426th position and valine at the 555th position in VP2. To develop a vaccine against CPV infection, we passaged the isolate 4 times in A72 cells.
RESULTS
The attenuated isolate conferred complete protection against lethal homologous CPV infection in dogs such that they did not develop any clinical symptoms, and their antibody titers against CPV were significantly high at 7-11 days post infection.
CONCLUSION
These results suggest that the virus isolate obtained after passaging can be developed as a novel vaccine against paroviral infection.

Keyword

Canine parvoviral infection; Vaccines; Novel type 2b; Dogs; Clinical isolate

MeSH Terms

Animals
Asparagine
Dogs
Enteritis
Parvovirus, Canine
Sequence Analysis, Protein
Sprains and Strains
Vaccination
Vaccines
Valine
Viruses
Asparagine
Vaccines
Valine

Figure

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  Fig. 1 Identification of canine parvovirus (CPV) in fecal sample from a dog with enteritis. Polymerase chain reaction was performed with the universal primer pairs (A) and CPV-2b-specific primer pairs (B) to identify CPV in the fecal sample ofa dog with enteritis. (A) M, marker; S1 and S2, fecal samples from a dog. (B) M1, 100 bp marker; M2, marker; S, fecal sample.

• 

  Fig. 2 Immunofluorescent assay (IFA) to verify the presence of the canine parvovirus (CPV). A72 cells were infected with the clinical isolate for 3 days, and the presence of CPV on infected cells was confirmed by IFA using anti-CPV monoclonal antibody and fluorescein isothiocyanate-conjugated anti-mouse IgG.

• 

  Fig. 3 Polymerase chain reaction (PCR) tests for detection of viremia and canine parvovirus (CPV) shedding. The (A) blood and (B) feces of dogs in the 3 different groups (group I, #1 and #2; group II, #3 and #4; group III, #5, #6, and #7) were taken at 1, 3, 5, 7, 9, and 11 days after inoculation and were subjected to PCR analysis to detect CPV.

• 

  Fig. 4 Changes in levels of antibody titers against canine parvovirus (CPV). Antibody titer levels against CPV at different times (days 0, 1, 3, 6, 7, 9, and 11 after inoculation) were determined by using ImmunoComb. The dogs in group II died at 6 dpi.

• 

  Fig. 5 Canine parvovirus (CPV) detection in different anatomical samples of the dogs after inoculation of the virus. On the day of death or 11 days after inoculation, 10 different anatomical samples from each dog were screened for the virus by polymerase chain reaction. Jej, jejunum; mLN, mesenteric lymph nodes; Thy, thymus; Lun, lung; Spl, spleen; Ile, ileum; Duo, duodenum; cMu, cardiac muscle; Liv, liver; Kid, kidney.

• 

  Fig. 6 Histopathological changes in the small intestine of dogs in group II. Hematoxylin and eosin staining was performed on the samples from duodenum and jejunum of the group II dogs that showed typical clinical signs of canine parvovirus infection. Histopathological changes in 2 dogs (dogs #3 and #4) were similar, and representative changes are shown here.

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