In this study, both the P. ovale wallikeri and P. knowlesi real-time PCR assays evaluated were proven to be very specific since no cross-reactivity was observed with the DNA obtained from the positive 128 samples containing other Plasmodium species, with the DNA obtained from blood protozoa other than Plasmodium, and with the DNA from the 270 negative samples for malaria (the veracity of which was guaranteed by the positivity of the β-actin internal control system and the negative results by the genus-specific PCR assays). The gene discussed is ACTB; the disease is malaria.