1921
Volume 39, Issue 1
  • ISSN: 0002-9637
  • E-ISSN: 1476-1645
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Abstract

Abstract

Blood specimens to be tested for the presence of using specific DNA probes can be stored as high salt lysates (HSL) without refrigeration. The lysates are prepared from 100 µl blood samples by a simple 3-step procedure using 2 volumes of HO to lyse the erythrocytes (step I), 1 volume of a detergent/EDTA mix to lyse the parasites (step II), followed by the addition of 1 volume cesium trifluoroacetate (CsTFA) (step III). The parasite DNA was found to be undegraded, as shown by the unaltered pattern of repetitive sequences obtained after storage of up to 1 month at 37°C, due to the inhibition of DNA degrading enzymes by the cesium salt. The bulk of protein can be removed from the samples by a 1-step precipitation. The addition of 0.3 volumes of a mixture of ethanol: chloroform: isoamyl alcohol (2.5:1:0.04 v/v) precipitates >90% of the proteins from the lysates, leaving >86% of the parasite DNA in the supernatant. The reduced protein content of the samples, when applied to solid supports, results in an increased signal: background ratio on autoradiograms.

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/content/journals/10.4269/ajtmh.1988.39.33
1988-07-01
2017-11-24
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http://instance.metastore.ingenta.com/content/journals/10.4269/ajtmh.1988.39.33
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  • Accepted : 16 Dec 1987

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