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The use of biotin-labelled, synthetic DNA oligomers for the detection and identification of Plasmodium falciparum

Published online by Cambridge University Press:  06 April 2009

M. A. Hughes
Affiliation:
Wolfson Unit of Molecular Genetics, Pembroke Place, Liverpool L3 SQA
M. Hommel
Affiliation:
Department of Tropical Medicine and Infectious Diseases, Liverpool School of Tropical Medicine, Pembroke Place, Liverpool L3 SQA
J. M. Crampton
Affiliation:
Wolfson Unit of Molecular Genetics, Pembroke Place, Liverpool L3 SQA

Summary

An oligonucleotide mixture based on the 21 base pair repeat sequence of Plasmodium falciparum was covalently coupled to biotin and used as a probe to detect P. falciparum DNA. The limit of detection was 10 ng. This method was further developed as a fingerprint assay for parasite strain typing. After restrictian enzyme digestion, blotting and hybridization, distinct banding patterns were obtained for the strains tested and these were reproducible. In addition, discrete differences were found between PLF-3 S + /S-strains which may implicate genetic reorganization in the switching mechanism which occurs when parasites are passed from an intact to a splenectomized animal.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1990

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