Novel gene targets are needed in accurate 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 the development of PCR assay for the detection of Plasmodium based on the (dlc-Tctex) as a genus and species-specific tool in malaria diagnosis. Multiple primers were designed based on Plasmodium spp dlc(Tctex) genes. The primers were applied on PCR to detect malaria on clinical samples and on laboratory maintained isolates of P. falciparum and P. vivax for human infecting species and P. knowlesi and P. cynomolgi for zoonoses infection involving primates. The amplified PCR fragments were gene cleaned and sequenced. BLASTn e-values output from the raw nucleotide queries supports that the genes are uniquely conserved. Species-specific primers amplified P. falciparum infections with no cross-reactivity to P. vivax, 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. Primers designed for Plasmodium genus amplified the target band in all clinical malaria samples but also had another specific band amplification. This preliminary 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 to differentiate mixed malaria infections.
The black rhino sanctuaries system has played a key role in repopulating and starting new subpopulations in Kenya. If this system is efficiently managed it may save the black rhinoceros from local extinction. Understanding the genetic status of endangered species is the most elemental sine qua non of animal breeding and conservation. It is therefore important to determine the genetic diversity of black rhino populations, especially of nucleus breeding populations that are used as a source of individuals for translocation and supplementation programs. We assessed the genetic diversity of one of the pioneer breeding subpopulations of the black rhino Diceros bicornis in Kenya using a mitochondrial DNA D-loop region. We then compared this subpopulation with the entire Kenyan ©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 18 (2): gmr18203 D.N. Thuo et al. 2 population to determine its status vis-à-vis the Kenya pooled population and determine the possible sources/relationships of the founder individuals. In the 469-bp D-loop region we analyzed, 7.11% of the sites were variants, contributing to 18 distinct haplotypes. Estimates of genetic diversity using haplotype and nucleotide diversity metrics showed that the Lake Nakuru National Park subpopulation has a slightly lower genetic diversity when compared with that of the pooled Kenya population. The phylogenetic tree revealed that Lake Nakuru National Park founder individuals were probably sourced from multiple subpopulations. The dendrogram and the principal coordinate analysis plot indicated that the Maasai Mara subpopulation is not a distinct subpopulation, as had been suggested previously. Our results provide baseline genetic data for the Lake Nakuru National Park breeding subpopulation and valuable information for translocation/supplementation programs.
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