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Leishmania braziliensis:Characterisation of a Complex Specific Subtelomeric Repeat Sequence and Its Use in the Detection of Parasites
Authors:Guoliang Fu  Georgia Perona-Wright  Douglas C. Barker
Affiliation:MRC Outstation of NIMR, Molteno Laboratories, Department of Pathology, University of Cambridge, CB2 1QP, U.K.
Abstract:Fu, G., Perona-Wright, G., and Barker, D. C. 1998.Leishmania braziliensis: Characterisation of a complex specific subtelomeric repeat sequence and its use in the detection of parasites.Experimental Parasitology90, 236–243. A 1.6-kb tandem repeat sequence had previously been identified in the subtelomeric region of mini- and megabase chromosomes fromLeishmania braziliensis.Southern hybridisation was used to demonstrate that the repeat is complex specific. The sequence was characterised in strains representing four species of theL. braziliensiscomplex. This data allowed an assessment of the evolutionary relationship of the four species. PCR primers targeted to the repeat amplify only DNA from species of theL. braziliensiscomplex. Titration assays indicate that a minimum of 50 fg of parasite DNA can be detected by PCR alone. Southern hybridisation increases the limit of detection to 5 fg. Interspecies variation in the repeat sequence enabled restriction enzyme digestion of PCR products to distinguish individual species within theL. braziliensiscomplex.
Keywords:Abbreviations: Leishmania braziliensiscomplex   subtelomeric repeat   diagnosis   restriction enzyme digestion   bp, base pairs   ELISA, enzyme-linked immunosorbent assay   IFAT, immunofluorescence antibody testing   kDNA, kinetoplast deoxyribonucleic acid   PCR, polymerase chain reaction.
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