This study aimed at identifying the effect of vapocoolant spray and EMLA (eutectic mixture of local anesthetics) cream upon DPT (diphtheria-pertussis-tetanus)vaccineassociated injection pain in infants. A nonequivalent control group pretest-posttest design was used. The subjects were 49 infants, 19 of them for control group, 15 of them for vapocoolant group, and 15 infants for EMLA group. Pulse and oxygen saturation as pain indicators were measured before and after DPT vaccination. FLACC was also measured after vaccination. The data were collected between October 2009 and June 2010 and analyzed using SPSS WIN 20.0. EMLA group had significant a little changes in pulse (F =43.37, p <.001) and oxygen saturation (F =9.86, p=.003) compared to the control and vapocoolant group. But there was no difference in FLACC pain score among three groups. This results showed that EMLA cream is an effective agent for reducing DPT vaccination-associated pain. Therefore, EMLA cream can be used to reduce pain at public health centers and clinical settings.