A rapid DNA assay for the species-specific detection and quantification of Brugia in blood samples.
Identifieur interne : 00DA76 ( Main/Exploration ); précédent : 00DA75; suivant : 00DA77A rapid DNA assay for the species-specific detection and quantification of Brugia in blood samples.
Auteurs : C B Poole [États-Unis] ; S A WilliamsSource :
- Molecular and biochemical parasitology [ 0166-6851 ] ; 1990.
Descripteurs français
- KwdFr :
- ADN (analyse), Animaux, Brugia (génétique), Brugia (isolement et purification), Filariose lymphatique (diagnostic), Gerbillinae, Humains, Hybridation d'acides nucléiques, Microfilaria (génétique), Microfilaria (isolement et purification), Sang (parasitologie), Sondes d'ADN, Spécificité d'espèce, Séquences répétées d'acides nucléiques.
- MESH :
- analyse : ADN.
- diagnostic : Filariose lymphatique.
- génétique : Brugia, Microfilaria.
- isolement et purification : Brugia, Microfilaria.
- parasitologie : Sang.
- Animaux, Gerbillinae, Humains, Hybridation d'acides nucléiques, Sondes d'ADN, Spécificité d'espèce, Séquences répétées d'acides nucléiques.
English descriptors
- KwdEn :
- Animals, Blood (parasitology), Brugia (genetics), Brugia (isolation & purification), DNA (analysis), DNA Probes, Elephantiasis, Filarial (diagnosis), Gerbillinae, Humans, Microfilariae (genetics), Microfilariae (isolation & purification), Nucleic Acid Hybridization, Repetitive Sequences, Nucleic Acid, Species Specificity.
- MESH :
- chemical , analysis : DNA.
- diagnosis : Elephantiasis, Filarial.
- genetics : Brugia, Microfilariae.
- isolation & purification : Brugia, Microfilariae.
- parasitology : Blood.
- Animals, DNA Probes, Gerbillinae, Humans, Nucleic Acid Hybridization, Repetitive Sequences, Nucleic Acid, Species Specificity.
Abstract
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.
PubMed: 2348829
Affiliations:
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">A rapid DNA assay for the species-specific detection and quantification of Brugia in blood samples.</title><author><name sortKey="Poole, C B" sort="Poole, C B" uniqKey="Poole C" first="C B" last="Poole">C B Poole</name><affiliation wicri:level="2"><nlm:affiliation>Department of Biological Sciences, Smith College, Northampton, MA 01063.</nlm:affiliation><country xml:lang="fr">États-Unis</country><placeName><region type="state">Massachusetts</region></placeName><wicri:cityArea>Department of Biological Sciences, Smith College, Northampton</wicri:cityArea></affiliation></author><author><name sortKey="Williams, S A" sort="Williams, S A" uniqKey="Williams S" first="S A" last="Williams">S A Williams</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="1990">1990</date><idno type="RBID">pubmed:2348829</idno><idno type="pmid">2348829</idno><idno 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samples.</title><author><name sortKey="Poole, C B" sort="Poole, C B" uniqKey="Poole C" first="C B" last="Poole">C B Poole</name><affiliation wicri:level="2"><nlm:affiliation>Department of Biological Sciences, Smith College, Northampton, MA 01063.</nlm:affiliation><country xml:lang="fr">États-Unis</country><placeName><region type="state">Massachusetts</region></placeName><wicri:cityArea>Department of Biological Sciences, Smith College, Northampton</wicri:cityArea></affiliation></author><author><name sortKey="Williams, S A" sort="Williams, S A" uniqKey="Williams S" first="S A" last="Williams">S A Williams</name></author></analytic><series><title level="j">Molecular and biochemical parasitology</title><idno type="ISSN">0166-6851</idno><imprint><date when="1990" type="published">1990</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Blood (parasitology)</term><term>Brugia (genetics)</term><term>Brugia (isolation & purification)</term><term>DNA (analysis)</term><term>DNA Probes</term><term>Elephantiasis, Filarial (diagnosis)</term><term>Gerbillinae</term><term>Humans</term><term>Microfilariae (genetics)</term><term>Microfilariae (isolation & purification)</term><term>Nucleic Acid Hybridization</term><term>Repetitive Sequences, Nucleic Acid</term><term>Species Specificity</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ADN (analyse)</term><term>Animaux</term><term>Brugia (génétique)</term><term>Brugia (isolement et purification)</term><term>Filariose lymphatique (diagnostic)</term><term>Gerbillinae</term><term>Humains</term><term>Hybridation d'acides nucléiques</term><term>Microfilaria (génétique)</term><term>Microfilaria (isolement et purification)</term><term>Sang (parasitologie)</term><term>Sondes d'ADN</term><term>Spécificité d'espèce</term><term>Séquences répétées d'acides nucléiques</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>DNA</term></keywords><keywords scheme="MESH" qualifier="analyse" xml:lang="fr"><term>ADN</term></keywords><keywords scheme="MESH" qualifier="diagnosis" xml:lang="en"><term>Elephantiasis, Filarial</term></keywords><keywords scheme="MESH" qualifier="diagnostic" xml:lang="fr"><term>Filariose lymphatique</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Brugia</term><term>Microfilariae</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Brugia</term><term>Microfilaria</term></keywords><keywords scheme="MESH" qualifier="isolation & purification" xml:lang="en"><term>Brugia</term><term>Microfilariae</term></keywords><keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Brugia</term><term>Microfilaria</term></keywords><keywords scheme="MESH" qualifier="parasitologie" xml:lang="fr"><term>Sang</term></keywords><keywords scheme="MESH" qualifier="parasitology" xml:lang="en"><term>Blood</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>DNA Probes</term><term>Gerbillinae</term><term>Humans</term><term>Nucleic Acid Hybridization</term><term>Repetitive Sequences, Nucleic Acid</term><term>Species Specificity</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Animaux</term><term>Gerbillinae</term><term>Humains</term><term>Hybridation d'acides nucléiques</term><term>Sondes d'ADN</term><term>Spécificité d'espèce</term><term>Séquences répétées d'acides nucléiques</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">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.</div></front></TEI><affiliations><list><country><li>États-Unis</li></country><region><li>Massachusetts</li></region></list><tree><noCountry><name sortKey="Williams, S A" sort="Williams, S A" uniqKey="Williams S" first="S A" last="Williams">S A Williams</name></noCountry><country name="États-Unis"><region name="Massachusetts"><name sortKey="Poole, C B" sort="Poole, C B" uniqKey="Poole C" first="C B" last="Poole">C B Poole</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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