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Use of a DNA Hybridization Assay for the Detection of Plasmodium Falciparumin Field Trials

Martin Holmberg* Department of Immunology, Karolinska Institute, Stockholm, Sweden
† Department of Medical Genetics, Uppsala University, Uppsala, Sweden

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Fiona C. Shenton‡ Medical Research Council Laboratories, Fajara, The Gambia

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Lena Franzén* Department of Immunology, Karolinska Institute, Stockholm, Sweden
† Department of Medical Genetics, Uppsala University, Uppsala, Sweden

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Kebba Janneh‡ Medical Research Council Laboratories, Fajara, The Gambia

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Robert W. Snow‡ Medical Research Council Laboratories, Fajara, The Gambia

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Ulf Pettersson† Department of Medical Genetics, Uppsala University, Uppsala, Sweden

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Hans Wigzell* Department of Immunology, Karolinska Institute, Stockholm, Sweden

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Brian M. Greenwood‡ Medical Research Council Laboratories, Fajara, The Gambia

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A DNA probe consisting of 21 base pair repeats obtained from a Tanzanian isolate of Plasmodium falciparum, cloned in pBR322 and labeled with 32P by nick translation was used to detect malaria parasitemia in samples obtained during a malaria survey undertaken in The Gambia. In an initial trial the hybridization assay had a specificity for P. falciparum of 100% and a sensitivity of 68%. False negative results were obtained only on samples with low parasitemia. Assay of red cells collected during an earlier malaria survey which had been stored for 1 year at ™20°C gave a higher level of sensitivity (85%), suggesting a beneficial effect from freezing and thawing. This was confirmed by examining in the same assay red cells processed immediately after collection and after 2 weeks of storage at ™20°C. Freezing and thawing gave a 21% increase in positivity, and a sensitivity of 100% was achieved with the frozen samples. Quantitation of autoradiographs by visual inspection and by scintillation counting gave a reasonable correlation with parasite counts. The DNA hybridization assay has considerable promise as an epidemiological tool.

Author Notes

Address reprint requests to: Martin Holmberg, Department of Medical Genetics, Box 589, Biomedical Centre, S-751 23 Uppsala, Sweden.
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