Medicine
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This repository contains the published and unpublished research of the Faculty of Medicine by the staff members of the faculty
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Item A Simplified non radioactive DNA probe technique for the field detection of sibling species A of the Anopheles culicifacies (Diptera: Culicidae) complex(University of Sri Jayewardenepura, 2010) de Silva, B.G.D.N.K.; Hapugoda, G.P.G.M.D.; Karunanayake, E.H.Three cloned highly repetitive DNA sequences Rp36, Rp217 and Rp234 isolated from An. culicifacies Giles, sensu lato were developed as non radioactive DNA probes by using a biotinylated labeling and colorimetric detection system. These non radioactive DNA probes distinguish sibling species A from species B and C of the An. culicifacies complex in a dot blot hybridization assay using single mosquito DNA extracts diluted 50 fold. The biotinylated Rp217 probe was further assayed in a more simple procedure which involves the hybridization of blots prepared from squashed mosquito heads. This technique avoids the separate extraction of mosquito DNA and facilitates a number of samples to be processed rapidly while also allowing several field analyses to be carried out on one mosquito specimen.Item ITS-2 secondary structures and phylogeny of Anopheles culicifacies species(Biomedical Informatics Pub. Group, 2008) Dassanayake, R.S.; Gunawardene, Y.I.N.S.; de Silva, B.G.D.N.K.BACKGROUND: Second internal transcribed spacer (ITS2) has proven to contain useful biological information at higher taxonomic levels. OBJECTIVES: This study was carried out to unravel the biological information in the ITS2 region of An. culicifacies and the internal relationships between the five species of Anopheles culicifacies.METHODOLOGY: In achieving these objectives, twenty two ITS2 sequences (approximately 370bp) of An. culicifacies species were retrieved from GenBank and secondary structures were generated. For the refinement of the primary structures, i.e. nucleotide sequence of ITS2 sequences, generated secondary structures were used. The improved ITS2 primary structures sequences were then aligned and used for the construction of phylogenetic trees. RESULTS AND DISCUSSIONS: ITS2 secondary structures of culicifacies closely resembled near universal eukaryotes secondary structure and had three helices, and the structures of helix II and distal region of helix III of ITS2 of An. culicifacies were strikingly similar to those regions of other organisms strengthening possible involvement of these regions in rRNA biogenesis. Phylogenetic analysis of improved ITS2 sequences revealed two main clades one representing sibling B, C and E and A and D in the other. CONCLUSIONS: Near sequence identity of ITS2 regions of the members in a particular clade indicate that this region is undergoing parallel evolution to perform clade specific RNA biogenesis. The divergence of certain isolates of An. culicifacies from main clades in phylogenetic analyses suggests the possible existence of camouflaged sub-species within the complex of culicifacies. Using the fixed nucleotide differences, we estimate that these two clades have diverged nearly 3.3 million years ago, while the sibling species in clade 2 are under less evolutionary pressure, which may have evolved much later than the members in clade 1.Item Seasonal Trends of Anopheles Culicifacies population at Gomadiyagala, a village in a North Western Province of Sri Lanka(University of Sri Jayewardenepura, 1998) de Silva, B.G.D.N.K.; Gunasekera, M.B.; Abeyewickreme, W.; Wickremasinghe, M.B.Item Screening of Anopheles crlicifacies population of Sri Lanka for sibling species A.(Thacker's Press and Directories for Indian Research Fund Association, 1998) de Silva, B.G.D.N.K.; Gunasekera, M.B.; Abeyewickreme, W.; Abhayawardena, T.A.; Karunanayake, E.H.A total of 1119 Anopheles culicifacies mosquitoes collected from various malaria endemic regions in Sri Lanka were examined using two DNA probes Rp217 and Rp234, which enable the differentiation of sibling species A from B and C species of An. culicifacies. Sibling species A was found to be absent.Item Development of DNA probes for the identification of sibling species A of the Anopheles culicifacies(Diptera Culicidae) Complex(CABI Publishing, 1995) Gunasekera, M.B.; de Silva, B.G.D.N.K.; Abeyewickreme, W.; Subbarao, S.K.; Nandadasa, H.G.; Karunanayake, E.H.Three highly repetitive DNA sequences, Rp36, Rp217 and Rp234, were isolated from A. culicifacies s.l. The cloned DNA sequences were found at a higher copy number in species B and C, than in species A of the A. culicifacies complex. These sequences may therefore be used as DNA probes to distinguish species A from the other 2 species, using a 200-fold dilution of a single mosquito DNA extract in a dot-blot hybridization assay. Rp36 and Rp217 were completely sequenced. Internal repeats were absent in Rp36. Two related core sequences of 134 and 16 bp were found tandemly repeated in Rp217. These probes enable the rapid detection of species A of A. culicifacies in field investigationsItem Colonization and characterization of Anopheles culicifacies Giles, the major malaria vector in Sri Lanka(Museum and Reference Centre, SEAMEO-TROPMED National Centre of Thailand, 1993) de Silva, B.G.D.N.K.; Ratnayake, W.E.; Gunasekera, M.B.; Abeyewickreme, W.; Karunanayake, E.H.; Wickremasinghe, M.B.