Maureen W. Kariuki, Elijah K.Githui, Andrew G. McArthur, Esther N. Magiri, Nyamu M. Njagi and Agatha C. Mwangemi
Molecular Genetics Laboratory, National Museums of Kenya, P.O Box 40658, Nairobi, Kenya
Novel technologies are needed in rapid diagnosis of malaria. Previous studies show that the dynein light chains(dlc) in Plasmodium are uniquely conserved within the species, possibly due to their role as the cargo adptor moiety. This study aimed at development of PCR assay for detection of Plasmodium based on the (dlc) as a genus and species-specific tool in malaria diagnosis. Multiple primers were designed based on Plasmodium spp dlc(Tctex) genes. The primers were then tested by PCR on malaria clinical samples and laboratory maintained isolates of P. falciparum, for human infecting species and P.knowlesi and P.cynomolg, for zoonosis infection involving primates. The target PCR fragments were gene cleaned, sequenced. BLASTn e-values support that the genes are uniquely conserved. Species-specific primers amplified P. falciparum infections and no cross-reactivity with P.knowlesi or P. cynomolgi species. In this assay only 11 out of the 30 microscope positive malaria positive clinical blood samples were positive for PCR detection of P. falciparum infection. The Plasmodium genus primers amplified the target band but also other non-specific bands. This data demonstrate that a species-specific dlc(Tctex) PCR assay can be used for detection of P. falciparum and optimized genus primers can be applied in mixed malaria infections.
Keyword: Plasmodium detection, dlc-Tctex, PCR.