불활화 참돔이리도바이러스병(RSIVD) 백신의 효능 평가를 위한 MN50 및 ROC 분석 기반 면역 보호 기준 설정
Establishment of MN₅₀-based immune protection threshold using ROC analysis for efficacy evaluation of inactivated RSIVD vaccine
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Quantitative evaluation of immune responses is essential for establishing reliable criteria to assess vaccine-induced protection in fish. In this study, the microneutralization titer (MN₅₀) was investigated as a potential functional indicator associated with vaccine-induced immunity in rock bream (Oplegnathus fasciatus) through the integration of serological analysis and receiver operating characteristic (ROC) evaluation. Fish were intraperitoneally immunized with three antigen concentrations (10^10, 10^9 , and 10^8 viral genome copies/mL) of a formalin-inactivated red sea bream iridovirus (RSIV) vac- cine, and both relative percent survival (RPS) and serum MN₅₀ titers were evaluated. Following the RSIV challenge, the vaccinated groups exhibited significantly higher survival than the control (p < 0.01), with RPS values of 90.0%, 66.7%, and 33.3% for the high-, middle-, and low-dose groups, respectively. The log₂ MN₅₀ antibody titers increased significantly from one week post-vaccination and remained consistently higher than those of the control group (3.01 ± 0.0), in which no neutralizing activity was detected. To quantitatively evaluate the association between MN₅₀ levels and protective efficacy, ROC curve analysis was performed among the vaccinated groups. The ROC analysis yielded an AUC of 0.7177 (95% CI: 0.6245–0.8108, p < 0.0001), and the optimal cut-off value calculated using Youden’s J index was log₂ MN50 > 8.489. These findings indicate that MN50 has potential applic-ability as a serological parameter associated with vaccine-induced protection in fish and may contribute to the development of standardized serological criteria for assessing aquaculture vaccines.
Abstract
Quantitative evaluation of immune responses is essential for establishing reliable criteria to assess vaccine-induced protection in fish. In this study, the microneutralization titer (MN₅₀) was investigated as a potential functional indicator associated with vaccine-induced immunity in rock bream (Oplegnathus fasciatus) through the integration of serological analysis and receiver operating characteristic (ROC) evaluation. Fish were intraperitoneally immunized with three antigen concentrations (10^10, 10^9 , and 10^8 viral genome copies/mL) of a formalin-inactivated red sea bream iridovirus (RSIV) vac- cine, and both relative percent survival (RPS) and serum MN₅₀ titers were evaluated. Following the RSIV challenge, the vaccinated groups exhibited significantly higher survival than the control (p < 0.01), with RPS values of 90.0%, 66.7%, and 33.3% for the high-, middle-, and low-dose groups, respectively. The log₂ MN₅₀ antibody titers increased significantly from one week post-vaccination and remained consistently higher than those of the control group (3.01 ± 0.0), in which no neutralizing activity was detected. To quantitatively evaluate the association between MN₅₀ levels and protective efficacy, ROC curve analysis was performed among the vaccinated groups. The ROC analysis yielded an AUC of 0.7177 (95% CI: 0.6245–0.8108, p < 0.0001), and the optimal cut-off value calculated using Youden’s J index was log₂ MN50 > 8.489. These findings indicate that MN50 has potential applic-ability as a serological parameter associated with vaccine-induced protection in fish and may contribute to the development of standardized serological criteria for assessing aquaculture vaccines.