Canine parvovirus-2 (CPV-2) has been reported worldwide as a major pathogen associated with acute hemorrhagic enteritis. The disease is a major infectious cause of death, particularly in young dogs. The earliest type of CPV-2 was replaced with three main subspecies, CPV-2a, CPV-2b, and CPV-2c, within a few years. Vaccination is carried out regularly, but the emergence of antigenic variants and the influence of maternal antibodies have limited the efficacy of commercial vaccines. New vaccines, such as the subunit vaccine, have been developed for alternative, safe, and effective vaccination. The baculovirus expression vector system (BEVS) is an excellent eukaryotic expression system with a high-level expression of foreign proteins and the ability of post-translational modification. Therefore, it is used widely to produce recombinant protein and subunit vaccines. In this study, the VP2 protein of CPV-2b cloned in the gateway vector system was generated using a baculovirus expression system in Spodoptera frugiperda (SF9) insect cells. Hemagglutination assay (HA) titers (24) were obtained, and the expression was detected in 6-His tagged VP2 and monoclonal antibody (mAb) against CPV-2 by western blotting. The VP2 protein of CPV-2b expressed in this study may provide a basis for a clinical diagnosis and vaccination applications for CPV-2.

Canine parvovirus type-2 (CPV-2) is a major etiological agent causing gastrointestinal enteritis in domestic and wild carnivores. Since the emergence of CPV-2 in the late 1970s, subtypes CPV-2a, CPV-2b, and CPV-2c have spread worldwide. CPV-2 prevalence differed according to region and season. This study aims to investigate the prevalence of CPV-2 infection in Korea. Samples were collected from 536 dog feces in animal shelters and 225 necropsied intestinal tissues of dog carcasses submitted in the Animal and Plant Quarantine Agency (APQA) for diagnostic purposes from 2016 to 2020 in Korea. Among the 761 samples, 181 (23.8%) were positive for the following subtypes: CPV-2a (n=138), CPV-2c (n=16), CPV-2b (n=14), and CPV-2 (n=2). Feline parvovirus (n=2) and co-infection with CPV-2a and CPV-2c (n=1) were also detected. There was no significant difference in the regional distribution of CPV-2 in Korea, which is prevalent in winter. This result shows the prevalence of CPV-2 according to various environments in Korea and will be useful in establishing an effective prevention strategy against CPV-2 that reflects the situation in Korea with continuous monitoring.

The canine parvovirus (CPV) causes clinical signs, such as severe enteritis, dehydration, diarrhea, vomiting, leukopenia, and hair loss, which may lead to death. Vaccination is still the most important approach, as no specific treatment exists to prevent CPV. Monoclonal antibodies are valuable tools to study the pathogenic mechanisms of CPV and develop effective diagnostic reagents and pharmaceuticals. In this study, two monoclonal antibodies (MAbs) against CPV-2a were obtained through hybridoma technology by fusing myeloma cells and B cells from the spleens of mice immunized with CPV type 2a (CPV-2a). Two MAbs (CPV-330 and CPV-620) were studied on the reactivity of vaccine (CPV-2a) and field strains (CPV-new 2a, -2b, and -2c) by indirect immunofluorescence (IFA), hemagglutination inhibition test (HI), virus neutralization test (VN), and inhibition of virus growth test. Two MAbs showed similar antibody titers for HI and VN. On the other hand, CPV-330 inhibited the viral replication in Crandell-Rees Feline Kidney (CRFK) cells better than CPV-620. These CPV MAbs may provide valuable biological reagents to study the CPV pathogenic mechanisms and work as therapeutic antibodies.

Canine parvovirus (CPV) remains a leading infectious cause of death in canines, especially in young puppies. Though vaccination is being carried out regularly, immunization failures occur, and puppies may be exposed to infection. Virus-like particles (VLPs) act like a subunit vaccine, mimicking the structure of authentic viruses. Therefore, VLPs have the potential to be used as vaccine candidates. Since Viral Protein 2 (VP2), a major structural protein of CPV, is the crucial antigen for CPV, the purpose of this study was to produce a recombinant VP2 of new canine parvovirus-2a using the baculovirus expression system in SF9 insect cells. The results revealed that recombinant VP2 assembles to form VLPs with antigenic properties similar to those of natural CPV, the recombinant VLP can produce a hemagglutination assay (HA) titer (1:210) in SF9 cells. Expression of the recombinant 6-His-tagged VP2 in SF9 cells was confirmed by western blotting. These findings suggest that the recombinant VP2 expressed in this study could be used as an efficient subunit vaccine against CPV infection.

Viral protein 2 (VP2), which is the structural protein of parvovirus, can produce virus-like particles (VLPs) by a self-assembly process in vitro, making VLPs attractive vaccine candidates. VP2 of canine parvovirus (CPV) is responsible for neutralizing antibodies in immunized animals. In this study, VP2 protein of canine parvovirus-2c was expressed using a baculovirus expression system and assembled into parvovirus-like particles in insect cells. The results show that VP2 proteins assembled into virus-like particles (VLPs) with antigenic properties similar to those of natural CPV and a high hemagglutination (HA) titer (1:27). The recombinant 6-His-tagged VP2 protein with a molecular mass of about 65 kDa was detected by anti-His antibody and anti-PPV serum. This study provides a foundation for the application of VP2 protein in the clinical diagnosis of CPV and in the vaccination against CPV.

Canine parvovirus type-2 (CPV-2) has been reported worldwide as the main agent associated with acute hemorrhagic enteritis, resulting in high morbidity, especially in young dogs. CPV-2 has three genetic variants, 2a, 2b, and 2c. Here, we report three cases of canine parvovirus enteritis associated with CPV-2a (2 samples) and -2c (1 sample) infections that occurred in three young dogs suffering from enteritis. Isolates from dog diarrheic fecal samples were sequenced by polymerase chain reaction (PCR) and identified as two types of CPV-2a and one type of CPV-2c. This work constitutes the first isolation and genetic characterization of CPV-2c in the Republic of Korea.

Canine parvovirus (CPV), a member of the genus Parvovirus, family Parvoviridae, is a significant causative agent in Canine reproductive failure, causing serious economic losses in the pet industry. The major capsid protein, VP2 is the main target protein for neutralizing antibodies in CPV. When VP2 was expressed using baculovirus, it was produced abundantly and assembled into virus-like particles (VLPs) similar in size and morphology to the original virions. It was named as CPV-VLP. Additionally, p35 sequences of canine distemper virus (CDV) as T-helper epitope were fusion-expressed with each of N-term, C-term or both sides of CPV-VP2. Production of double antigenic recombinant protein and formation of VLPs were analyzed by SDS-PAGE and transmission electron microscopy, respectively.

Canine parvovirus (CPV2) is one of the most virulent virus causing acute hemorrhagic enteritis and myocarditis in dogs. Infection mainly caused by the ingestion of virus through the mucosal route. Therefore, induction of mucosal immunity is essential in prevention of Canine Parvovirus (CPV2) infection. For safe and effective delivery of viral antigens to the mucosal immune system, a novel surface antigen display system for lactic acid bacteria using the poly-γ-glutamic acid synthetase A protein (pgsA) of Bacillus subtilis as an anchoring matrix was applied in order to display CPV2 antigen on the surface of the recombinant L. casei. Recombinant fusion proteins comprised of pgsA and the capsid protein (VP2-S1) showed stable expression in Lactobacillus casei. Surface localization of the fusion protein was verified by cellular fractionation analyses. Oral and nasal inoculations of recombinant L. casei into mice resulted in high levels of serum immunoglobulin G (IgG) and mucosal IgA, as demonstrated by ELISA using recombinant VP2-S1 proteins. Mice receiving intranasal immunization mounted higher antibody response than those receiving oral immunization. These results indicate that mucosal immunization with recombinant L. casei expressing CPV2 VP2-S1 protein on its surface provides an effective means for elicitation of strong antibody responses against CPV 2 VP2-S1.

Canine parvovirus (CPV) type 2a (CPV-2a) has recently been identified as the main genotype circulating in the dog population in South Korea. Although CPV vaccines protect domestic dogs from CPV-2 infection, the efficacy of commercial live or inactivated CPV vaccines against CPV-2a has not been reevaluated. In this study, dogs were immunized with one of 7 commercial CPV vaccines (4 modified live and 3 inactivated vaccines) followed by challenge with CPV-2a strain, KV0901 that had been isolated from naturally infected dog in 2009. All dogs vaccinated twice with 4 commercial modified live CPV vaccines were seroconverted (geometric mean HI titer > 190.2) and most of dogs were completely protected against virulent CPV-2a strain infection. The dogs inoculated with 3 commercial inactivated CPV vaccines were also seroconverted and showed a slight loss of appetite and light diarrhea for 4 days after challenge and returned to normal at 5 days post challenge. However, the non-vaccinated dogs revealed the typical clinical signs of CPV infection including haemorrhgic diarrhea. In conclusion, the 4 live CPV vaccines licensed in Korea cross-protected dogs against virulent challenge with CPV-2a and are applicalble to pet dogs for the prevention of CPV infection.