A polymerase chain reaction assay for detection of the parasite Wuchereria bancrofti in human blood samples.
Identifieur interne : 00C596 ( Main/Merge ); précédent : 00C595; suivant : 00C597A polymerase chain reaction assay for detection of the parasite Wuchereria bancrofti in human blood samples.
Auteurs : M. Zhong [États-Unis] ; J. Mccarthy ; L. Bierwert ; M. Lizotte-Waniewski ; S. Chanteau ; T B Nutman ; E A Ottesen ; S A WilliamsSource :
- The American journal of tropical medicine and hygiene [ 0002-9637 ] ; 1996.
Descripteurs français
- KwdFr :
- ADN des helminthes (), ADN des helminthes (sang), Amorces ADN (), Animaux, Clonage moléculaire, Données de séquences moléculaires, Filariose lymphatique (diagnostic), Filariose lymphatique (parasitologie), Humains, Parasitémie (diagnostic), Parasitémie (parasitologie), Réaction de polymérisation en chaîne, Sensibilité et spécificité, Spécificité d'espèce, Séquence nucléotidique, Séquences répétées d'acides nucléiques, Wuchereria bancrofti (génétique), Wuchereria bancrofti (isolement et purification).
- MESH :
- diagnostic : Filariose lymphatique, Parasitémie.
- génétique : Wuchereria bancrofti.
- isolement et purification : Wuchereria bancrofti.
- parasitologie : Filariose lymphatique, Parasitémie.
- sang : ADN des helminthes.
- ADN des helminthes, Amorces ADN, Animaux, Clonage moléculaire, Données de séquences moléculaires, Humains, Réaction de polymérisation en chaîne, Sensibilité et spécificité, Spécificité d'espèce, Séquence nucléotidique, Séquences répétées d'acides nucléiques.
English descriptors
- KwdEn :
- Animals, Base Sequence, Cloning, Molecular, DNA Primers (chemistry), DNA, Helminth (blood), DNA, Helminth (chemistry), Elephantiasis, Filarial (diagnosis), Elephantiasis, Filarial (parasitology), Humans, Molecular Sequence Data, Parasitemia (diagnosis), Parasitemia (parasitology), Polymerase Chain Reaction, Repetitive Sequences, Nucleic Acid, Sensitivity and Specificity, Species Specificity, Wuchereria bancrofti (genetics), Wuchereria bancrofti (isolation & purification).
- MESH :
- chemical , blood : DNA, Helminth.
- chemical , chemistry : DNA Primers, DNA, Helminth.
- diagnosis : Elephantiasis, Filarial, Parasitemia.
- genetics : Wuchereria bancrofti.
- isolation & purification : Wuchereria bancrofti.
- parasitology : Elephantiasis, Filarial, Parasitemia.
- Animals, Base Sequence, Cloning, Molecular, Humans, Molecular Sequence Data, Polymerase Chain Reaction, Repetitive Sequences, Nucleic Acid, Sensitivity and Specificity, Species Specificity.
Abstract
To identify Wuchereria bancrofti DNA sequences that could be used as the basis for a simple and rapid parasite detection assay, a genomic library of W. bancrofti was constructed and screened for highly repeated DNA. The repeat found with the highest copy number was 195 basepairs (bps) long, 77% AT, and 300 copies per haploid genome. This sequence was designated the Ssp I repeat because it has a unique recognition site for that restriction endonuclease in all or most of the repeat copies. The Ssp I repeat DNA family is dispersed, genus-specific, and exists in all of the different geographic isolates of W. bancrofti tested. Based on DNA sequence analysis of this repeat, we have developed an assay to detect very small quantities of W. bancrofti DNA using the polymerase chain reaction (PCR). With this PCR assay, the Ssp I repeat was detected in as little as 1 pg of w. bancrofti genomic DNA (about 1% of the DNA in one microfilaria) added to 100 microliters of human blood. The PCR assay also amplified Ssp I repeat DNA from geographic isolates of W. bancrofti from around the world but not from other species of filariae or from human or mosquito DNA. Microfilaria-positive human blood samples collected in Mauke, Cook Islands were shown to be Ssp I PCR-positive, while microfilaria-negative samples were PCR-negative. The specificity and sensitivity of the Ssp I PCR assay indicates that this approach has significant potential for improved screening of large human populations for active W. bancrofti infection.
PubMed: 8615447
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pubmed:8615447Le document en format XML
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Bierwert</name></author><author><name sortKey="Lizotte Waniewski, M" sort="Lizotte Waniewski, M" uniqKey="Lizotte Waniewski M" first="M" last="Lizotte-Waniewski">M. Lizotte-Waniewski</name></author><author><name sortKey="Chanteau, S" sort="Chanteau, S" uniqKey="Chanteau S" first="S" last="Chanteau">S. Chanteau</name></author><author><name sortKey="Nutman, T B" sort="Nutman, T B" uniqKey="Nutman T" first="T B" last="Nutman">T B Nutman</name></author><author><name sortKey="Ottesen, E A" sort="Ottesen, E A" uniqKey="Ottesen E" first="E A" last="Ottesen">E A Ottesen</name></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="1996">1996</date><idno type="RBID">pubmed:8615447</idno><idno type="pmid">8615447</idno><idno type="wicri:Area/PubMed/Corpus">005198</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">005198</idno><idno type="wicri:Area/PubMed/Curation">005198</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">005198</idno><idno type="wicri:Area/PubMed/Checkpoint">005198</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">005198</idno><idno type="wicri:Area/Ncbi/Merge">00B688</idno><idno type="wicri:Area/Ncbi/Curation">00B688</idno><idno type="wicri:Area/Ncbi/Checkpoint">00B688</idno><idno type="wicri:doubleKey">0002-9637:1996:Zhong M:a:polymerase:chain</idno><idno type="wicri:Area/Main/Merge">00C596</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">A polymerase chain reaction assay for detection of the parasite Wuchereria bancrofti in human blood samples.</title><author><name sortKey="Zhong, M" sort="Zhong, M" uniqKey="Zhong M" first="M" last="Zhong">M. Zhong</name><affiliation wicri:level="1"><nlm:affiliation>Department of Biological Sciences, Clark Science Center, Smith College, Northampton, Massachusetts 01063, USA.</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Biological Sciences, Clark Science Center, Smith College, Northampton, Massachusetts 01063</wicri:regionArea><wicri:noRegion>Massachusetts 01063</wicri:noRegion></affiliation></author><author><name sortKey="Mccarthy, J" sort="Mccarthy, J" uniqKey="Mccarthy J" first="J" last="Mccarthy">J. Mccarthy</name></author><author><name sortKey="Bierwert, L" sort="Bierwert, L" uniqKey="Bierwert L" first="L" last="Bierwert">L. Bierwert</name></author><author><name sortKey="Lizotte Waniewski, M" sort="Lizotte Waniewski, M" uniqKey="Lizotte Waniewski M" first="M" last="Lizotte-Waniewski">M. Lizotte-Waniewski</name></author><author><name sortKey="Chanteau, S" sort="Chanteau, S" uniqKey="Chanteau S" first="S" last="Chanteau">S. Chanteau</name></author><author><name sortKey="Nutman, T B" sort="Nutman, T B" uniqKey="Nutman T" first="T B" last="Nutman">T B Nutman</name></author><author><name sortKey="Ottesen, E A" sort="Ottesen, E A" uniqKey="Ottesen E" first="E A" last="Ottesen">E A Ottesen</name></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">The American journal of tropical medicine and hygiene</title><idno type="ISSN">0002-9637</idno><imprint><date when="1996" type="published">1996</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Base Sequence</term><term>Cloning, Molecular</term><term>DNA Primers (chemistry)</term><term>DNA, Helminth (blood)</term><term>DNA, Helminth (chemistry)</term><term>Elephantiasis, Filarial (diagnosis)</term><term>Elephantiasis, Filarial (parasitology)</term><term>Humans</term><term>Molecular Sequence Data</term><term>Parasitemia (diagnosis)</term><term>Parasitemia (parasitology)</term><term>Polymerase Chain Reaction</term><term>Repetitive Sequences, Nucleic Acid</term><term>Sensitivity and Specificity</term><term>Species Specificity</term><term>Wuchereria bancrofti (genetics)</term><term>Wuchereria bancrofti (isolation & purification)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ADN des helminthes ()</term><term>ADN des helminthes (sang)</term><term>Amorces ADN ()</term><term>Animaux</term><term>Clonage moléculaire</term><term>Données de séquences moléculaires</term><term>Filariose lymphatique (diagnostic)</term><term>Filariose lymphatique (parasitologie)</term><term>Humains</term><term>Parasitémie (diagnostic)</term><term>Parasitémie (parasitologie)</term><term>Réaction de polymérisation en chaîne</term><term>Sensibilité et spécificité</term><term>Spécificité d'espèce</term><term>Séquence nucléotidique</term><term>Séquences répétées d'acides nucléiques</term><term>Wuchereria bancrofti (génétique)</term><term>Wuchereria bancrofti (isolement et purification)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="blood" xml:lang="en"><term>DNA, Helminth</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>DNA Primers</term><term>DNA, Helminth</term></keywords><keywords scheme="MESH" qualifier="diagnosis" xml:lang="en"><term>Elephantiasis, Filarial</term><term>Parasitemia</term></keywords><keywords scheme="MESH" qualifier="diagnostic" xml:lang="fr"><term>Filariose lymphatique</term><term>Parasitémie</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Wuchereria bancrofti</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Wuchereria bancrofti</term></keywords><keywords scheme="MESH" qualifier="isolation & purification" xml:lang="en"><term>Wuchereria bancrofti</term></keywords><keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Wuchereria bancrofti</term></keywords><keywords scheme="MESH" qualifier="parasitologie" xml:lang="fr"><term>Filariose lymphatique</term><term>Parasitémie</term></keywords><keywords scheme="MESH" qualifier="parasitology" xml:lang="en"><term>Elephantiasis, Filarial</term><term>Parasitemia</term></keywords><keywords scheme="MESH" qualifier="sang" xml:lang="fr"><term>ADN des helminthes</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Base Sequence</term><term>Cloning, Molecular</term><term>Humans</term><term>Molecular Sequence Data</term><term>Polymerase Chain Reaction</term><term>Repetitive Sequences, Nucleic Acid</term><term>Sensitivity and Specificity</term><term>Species Specificity</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>ADN des helminthes</term><term>Amorces ADN</term><term>Animaux</term><term>Clonage moléculaire</term><term>Données de séquences moléculaires</term><term>Humains</term><term>Réaction de polymérisation en chaîne</term><term>Sensibilité et spécificité</term><term>Spécificité d'espèce</term><term>Séquence nucléotidique</term><term>Séquences répétées d'acides nucléiques</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">To identify Wuchereria bancrofti DNA sequences that could be used as the basis for a simple and rapid parasite detection assay, a genomic library of W. bancrofti was constructed and screened for highly repeated DNA. The repeat found with the highest copy number was 195 basepairs (bps) long, 77% AT, and 300 copies per haploid genome. This sequence was designated the Ssp I repeat because it has a unique recognition site for that restriction endonuclease in all or most of the repeat copies. The Ssp I repeat DNA family is dispersed, genus-specific, and exists in all of the different geographic isolates of W. bancrofti tested. Based on DNA sequence analysis of this repeat, we have developed an assay to detect very small quantities of W. bancrofti DNA using the polymerase chain reaction (PCR). With this PCR assay, the Ssp I repeat was detected in as little as 1 pg of w. bancrofti genomic DNA (about 1% of the DNA in one microfilaria) added to 100 microliters of human blood. The PCR assay also amplified Ssp I repeat DNA from geographic isolates of W. bancrofti from around the world but not from other species of filariae or from human or mosquito DNA. Microfilaria-positive human blood samples collected in Mauke, Cook Islands were shown to be Ssp I PCR-positive, while microfilaria-negative samples were PCR-negative. The specificity and sensitivity of the Ssp I PCR assay indicates that this approach has significant potential for improved screening of large human populations for active W. bancrofti infection.</div></front></TEI></record>Pour manipuler ce document sous Unix (Dilib)
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