Developing new primers for molecular identification of plasmodium spp. recorded in Sabah

Malaria is a major public health problem in tropical and subtropical areas, caused by five species of Plasmodium (P. falciparum, P. vivax, P. malariae, P. ovale andP. knowlesi) and is the leading cause of morbidity and mortality worldwide. We have developed molecular markers for three genes viz, Cytb, dhfr and Msp-1 gene and designed a protocol for rapid molecular diagnostics of the four malaria parasites prevalent in Southeast Asia. The new primers were used on the blood samples containing Plasmodium parasites by conventional PCR. The result was compared with the nested PCR of Singh et al. (2004) and the microscopy method. The result shows that the new set of primers had successfully amplified all four human malaria parasite species. These primers were 100% sensitive and more specific than microscopy and PCR identification using these primers was faster than the nested PCR. These alternative primers should provide powerful and rapid molecular diagnostic method for detecting Plasmodium species as well as providing reliable data for epidemiology study. These primers have the potential to be combined and used in multiplex PCR