A new competitive enzyme-linked immunosorbent assay demonstrates adequate immune levels to rabies virus in compulsorily vaccinated Japanese domestic dogs

Abstract

A competitive enzyme-linked immunosorbent assay (c-ELISA) was developed as an alternative to the viral neutralization (VN) test for rapid and simple detection of antibodies to rabies virus. The competitor antibody in the c-ELISA was a biotinylated monoclonal antibody to the nucleoprotein of rabies virus. Initial comparisons showed a high correlation between titers obtained with the VN test and the c-ELISA (n ‫؍‬ 88, r ‫؍‬ 0.90), indicating that the c-ELISA could be used as a reliable substitute for the VN test. To evaluate the immune status of Japanese dogs to rabies virus, a total of 1,019 serum samples were collected from domestic dogs in 1994 and tested for antibodies with the c-ELISA. Overall, 84.8% of the dogs had antibodies against rabies virus, indicating that the vaccination strategy for preventing rabies outbreaks in domestic dogs is probably sufficient in Japan. Dogs receiving final vaccinations a year or more previously were 48.3 and 90.3% positive for antibodies when vaccinated once only or two or more times, respectively. This suggests that almost all dogs vaccinated twice or more remain seropositive for over 1 year in Japan.

Comparison of antibody responses after vaccination with two inactivated rabies vaccines

“…This early peak is consistent with results obtained in other studies on experimental dogs (Minke, personnel observation, Kallel et al 2006). Peak antibody response in pets is classically reported in the literature between 3 and 6 weeks after vaccination (Sugiyama et al 1997, National Association of State Public Health Veterinarians 2007, Barth et al 1985, but laboratory dogs are known to respond better to vaccination than pet dogs (Aubert, 1992). An in-depth review of many experimental studies (Aubert, 1993) has shown a strong correlation between the development of rabies antibodies after vaccination and protection against rabies infection.…”

Validation and Standardization of Virus Neutralizing Test Using Indirect Immunoperoxidase Technique for the Quantification of Antibodies to Rabies Virus

“…Then, this study showed that the VNT-IIP correlated the FAVNT (r = 0.92, n = 248), prescribed as well as the RFFIT. Although these previous data (Sugiyama et al, 1997;Shimazaki et al, 2003;Cardoso et al, 2004) were not directly analysed with this study on account of some modification of materials, these studies indicated that the indirect immunoperoxidase technique strongly correlated with the other assays including standard methods. Furthermore, this study showed the validation of the indirect immunoperoxidase assay for the first time.…”

“…There are some reports regarding to the indirect immunoperoxidase assay for rabies serological test. Sugiyama et al (1997) and Cardoso et al (2004) reported that the assay correlates the competitive ELISA (r = 0.90, n = 88) and the mouse neutralization test (r = 0.99, n = 100 and r = 0.93, n = 99) respectively. Shimazaki et al (2003) also reported that it correlated the RFFIT (r = 0.95, n = 135), the OIE-prescribed method.…”

“…In this study, we selected anti-rabies monoclonal antibody as well as Shimazaki et al (2003) while Sugiyama et al (1997) and Cardoso et al (2004) used polyclonal antibody. In general, monoclonal antibody has the advantages of high specificity, easy quality control and stable and safe production even though the cost for establishment of hybridoma is expensive.…”

“…Since diagnosticians can view the obtained results by the naked eye, it is easy to check a large number of microplates. Although there are some reports regarding the VNT-IIP (Hirayama et al, 1990;Sugiyama et al, 1997;Shimazaki et al, 2003;Cardoso et al, 2004), the validation of this method has not been described. The purpose of this study was to validate the VNT-IIP and compare it with one of the international standard methods, the FAVNT.…”