Typhoid fever, a serious systemic infection caused by Salmonella enterica serovar Typhi, breaks out in developing countries. However, existing vaccines only induce relatively low protective effects with humoral responses and do not stimulate secondary immune response, especially to young people. The objective of this study is to evaluate the immunogenicity of the vaccine containing virulence capsular polysaccharide (Vi) conjugated with the optimal ratios of non-toxic variant of diphtheria toxin (CRM(197)) in mice. Six-week-old BALB/c female mice were injected intraperitoneally three times at intervals of 14 days and sera were collected on days 0, 14, 28, 42 and 56 post-injection. The efficacy of the vaccine was evaluated by comparing between negative control group injected with PBS and vaccine groups injected with Vi or Vi-CRM(197) conjugate of different ratio. Vi and CRM(197)-specific antibody responses were evaluated using enzyme-linked immunosorbent assay. The result showed that Vi-CRM(197)-1 group revealed the highest and significant Vi-specific IgG immune responses among the other groups and Vi group (p < 0.01). In conclusion, Vi-CRM(197)-1 conjugate vaccine induced the highest humoral immune response in mice and may be used as an effective vaccine to replace the existing typhoid vaccine for infants under 2 years old.