Protein arrays are powerful tools for antibody profiling and vaccine development against infectious agents. In the previous report, we successfully applied an antibody-based protein array for immunoprofiling of Plasmodium vivax infection. Herein, we developed a Ni-NTA surface based protein array to detect immune responses against the recombinant C-terminal region (19 and 42 kDa) of the P. vivax merozoite surface protein 1 (PvMSP1-19 and -42) from sera of vivax malaria patients. The PvMSP1-19 arrays detected P. vivax in 112 of 130 (86.2%; 95% CI, 83.2-89.2%) microscopically positive samples and 2 false positives were obtained among 100 sera samples from healthy subjects (2.0%; 95% CI, 0.6-3.4%). These results were in concordance with results of enzyme-linked immunosorbent assays (ELISA). Kappa values represented excellent agreement for the recombinant PvMSP1-19 protein against sera samples as measured by protein arrays and ELISA (Kappa=0.904, 95% CI: 0.849-0.960). The PvMSP1-42 protein arrays detected antibody response in 100 of 130 microscopically positive samples (76.9%; 95% CI, 72.4-86.8%) and 8 false positives were obtained in 100 healthy subjects (8.0%; 95% CI, 2.7-13.3%). There is no significant difference between the fluorescent intensity of antibody response to PvMSP1-19 and PvMSP1-42 in the positive sera samples (P>0.05). The novel protein array platform may be used for profiling naturally acquired humoral immune responses to P. vivax infection.