Leishmania braziliensis:Characterisation of a Complex Specific Subtelomeric Repeat Sequence and Its Use in the Detection of Parasites |
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Authors: | Guoliang Fu Georgia Perona-Wright Douglas C. Barker |
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Affiliation: | MRC Outstation of NIMR, Molteno Laboratories, Department of Pathology, University of Cambridge, CB2 1QP, U.K. |
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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. |
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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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