1921
Volume 37, Issue 1
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645
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Abstract

Abstract

Synthetic DNA oligomers homologous to 21-base long repetitive sequences of DNA were labeled with P using T4 kinase, and were hybridized with purified DNA and with processed blood samples from Africa. The sequences PFR1, its antiparallel oligomer PFR1R, and PFR1 covalently attached to biotin hybridized similarly to DNA. One-microliter aliquots of blood from Zaire spotted on prewet nylon filters and hybridized with PFR1 gave detectable autoradiogram signals from samples with parasitemias as low as 1,000 parasites/mm. Blood lysis and protein digestion followed by alkylation allowed dot-blot processing of larger aliquots of blood. After hybridization with PFR 1 and autoradiography, 26 samples were scored positive visually, compared with 34 scored positive by microscopy. The effective sensitivity for processed 10-µl samples was about 500 parasites/mm. Signals from hybridized probes were quantitated by liquid scintillation counting and densitometry, and were proportional to the amounts of purified DNA applied to the filter. Autoradiogram signals also were roughly proportional (correlation coefficient, r = 0.77) to the number of parasites/mm of blood from field samples as determined by microscopic examination.

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/content/journals/10.4269/ajtmh.1987.37.27
1987-07-01
2017-09-19
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http://instance.metastore.ingenta.com/content/journals/10.4269/ajtmh.1987.37.27
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  • Accepted : 31 Dec 1986

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