A rapid DNA assay for the species-specific detection and quantification of Brugia in blood samples.
This report describes a new assay for detecting filarial parasites of the genus Brugia in blood samples using labeled DNA probes. The sequences of these DNA probes are based on the HhaI repeat DNA family found in the genus Brugia. These DNA probes are species-specific and can detect the DNA from a single microfilaria in hybridization assays. To adapt this test for use on blood samples collected in the field, complex steps to separate microfilariae from blood cells and to purify parasite DNA were eliminated. We found that the most effective method was to filter blood samples through 5.0 microns pore nitrocellulose membranes, lyse the microfilariae on the membranes by proteinase K digestion, denature the parasite DNA with sodium hydroxide, and hybridize with the DNA probe. With this method, individual microfilariae can be visualized and counted on autoradiograms. The assay was evaluated in a mock field study using Brugia malayi-infected jirds and was found to be an efficient and accurate method for quantifying microfilariae in blood samples.
Poole
C B
CB
Department of Biological Sciences, Smith College, Northampton, MA 01063.
Williams
S A
SA
D000818:Animals; D001769:Blood / Q000469:parasitology*; D002009:Brugia / Q000235:genetics / Q000302:isolation & purification*; D004247:DNA / Q000032:analysis*; D015342:DNA Probes; D004605:Elephantiasis, Filarial / Q000175:diagnosis; D005849:Gerbillinae; D006801:Humans; D008842:Microfilariae / Q000235:genetics / Q000302:isolation & purification; D009693:Nucleic Acid Hybridization; D012091:Repetitive Sequences, Nucleic Acid; D013045:Species Specificity
D015342:DNA Probes; D004247:DNA