Development of a quantitative, competitive polymerase chain reaction--enzyme-linked immunosorbent assay for the detection of Wuchereria bancrofti DNA.
Identifieur interne : 00AA26 ( Main/Exploration ); précédent : 00AA25; suivant : 00AA27Development of a quantitative, competitive polymerase chain reaction--enzyme-linked immunosorbent assay for the detection of Wuchereria bancrofti DNA.
Auteurs : P. Fischer [États-Unis] ; X. Liu ; M. Lizotte-Waniewski ; I H Kamal ; R M Ramzy ; S A WilliamsSource :
- Parasitology research [ 0932-0113 ] ; 1999.
Descripteurs français
- KwdFr :
- ADN des helminthes (génétique), ADN des helminthes (sang), Analyse de régression, Animaux, Données de séquences moléculaires, Filariose lymphatique (diagnostic), Filariose lymphatique (sang), Humains, Reproductibilité des résultats, Réaction de polymérisation en chaîne (), Sensibilité et spécificité, Sondes d'ADN, Séquence nucléotidique, Séquences répétées d'acides nucléiques, Test ELISA (), Type II site-specific deoxyribonuclease, Wuchereria bancrofti (génétique), Wuchereria bancrofti (isolement et purification).
- MESH :
- diagnostic : Filariose lymphatique.
- génétique : ADN des helminthes, Wuchereria bancrofti.
- isolement et purification : Wuchereria bancrofti.
- sang : ADN des helminthes, Filariose lymphatique.
- Analyse de régression, Animaux, Données de séquences moléculaires, Humains, Reproductibilité des résultats, Réaction de polymérisation en chaîne, Sensibilité et spécificité, Sondes d'ADN, Séquence nucléotidique, Séquences répétées d'acides nucléiques, Test ELISA, Type II site-specific deoxyribonuclease.
English descriptors
- KwdEn :
- Animals, Base Sequence, DNA Probes, DNA, Helminth (blood), DNA, Helminth (genetics), Deoxyribonucleases, Type II Site-Specific, Elephantiasis, Filarial (blood), Elephantiasis, Filarial (diagnosis), Enzyme-Linked Immunosorbent Assay (methods), Humans, Molecular Sequence Data, Polymerase Chain Reaction (methods), Regression Analysis, Repetitive Sequences, Nucleic Acid, Reproducibility of Results, Sensitivity and Specificity, Wuchereria bancrofti (genetics), Wuchereria bancrofti (isolation & purification).
- MESH :
- chemical , blood : DNA, Helminth.
- chemical , genetics : DNA, Helminth.
- chemical : DNA Probes, Deoxyribonucleases, Type II Site-Specific.
- blood : Elephantiasis, Filarial.
- diagnosis : Elephantiasis, Filarial.
- genetics : Wuchereria bancrofti.
- isolation & purification : Wuchereria bancrofti.
- methods : Enzyme-Linked Immunosorbent Assay, Polymerase Chain Reaction.
- Animals, Base Sequence, Humans, Molecular Sequence Data, Regression Analysis, Repetitive Sequences, Nucleic Acid, Reproducibility of Results, Sensitivity and Specificity.
Abstract
A quantitative, competitive polymerase chain reaction (QC-PCR) assay for the sensitive detection of Wuchereria bancrofti DNA was developed. A competitor sequence was constructed by an exchange of nucleotides in the Wuchereria-specific Ssp I repeat. The PCR products were hybridized to specific DNA probes and their amounts, determined by an enzyme-linked immunosorbent assay (ELISA). In laboratory-prepared samples the QC-PCR-ELISA assay was capable of detecting the amount of DNA equivalent to 0.1 microfilaria (mf) added to 200 microl of blood lysate. The assay was also tested on 78 blood samples collected in endemic areas in Egypt. All 28 samples that were positive both for mf and for circulating antigen were also QC-PCR-ELISA-positive. In addition, one mf-negative but antigen-positive sample was also positive as determined by QC-PCR-ELISA. A positive correlation of mf density with the QC-PCR-ELISA was observed. Samples containing 10 or fewer mf/ml had a mean relative amount of Ssp I PCR product of 19.7 units, whereas samples with 11-100 mf/ml had a mean of 36.3 units and those with more than 100 mf/ml had a mean of 84.6 units. Because of the high standard deviation within each group, estimates of worm burdens in infected individuals using the QC-PCR-ELISA are not recommended. However, we present data indicating that the W. bancrofti QC-PCR-ELISA is a powerful new tool for evaluation of parasitic loads for community-based diagnosis of bancroftian filariasis.
PubMed: 9951959
Affiliations:
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Fischer</name><affiliation wicri:level="2"><nlm:affiliation>Department of Biological Sciences, Smith College, Northampton, MA 01063, USA. Pfischer@sophia.smith.edu</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Department of Biological Sciences, Smith College, Northampton, MA 01063</wicri:regionArea><placeName><region type="state">Massachusetts</region></placeName></affiliation></author><author><name sortKey="Liu, X" sort="Liu, X" uniqKey="Liu X" first="X" last="Liu">X. Liu</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="Kamal, I H" sort="Kamal, I H" uniqKey="Kamal I" first="I H" last="Kamal">I H Kamal</name></author><author><name sortKey="Ramzy, R M" sort="Ramzy, R M" uniqKey="Ramzy R" first="R M" last="Ramzy">R M Ramzy</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">Parasitology research</title><idno type="ISSN">0932-0113</idno><imprint><date when="1999" type="published">1999</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Base Sequence</term><term>DNA Probes</term><term>DNA, Helminth (blood)</term><term>DNA, Helminth (genetics)</term><term>Deoxyribonucleases, Type II Site-Specific</term><term>Elephantiasis, Filarial (blood)</term><term>Elephantiasis, Filarial (diagnosis)</term><term>Enzyme-Linked Immunosorbent Assay (methods)</term><term>Humans</term><term>Molecular Sequence Data</term><term>Polymerase Chain Reaction (methods)</term><term>Regression Analysis</term><term>Repetitive Sequences, Nucleic Acid</term><term>Reproducibility of Results</term><term>Sensitivity and Specificity</term><term>Wuchereria bancrofti (genetics)</term><term>Wuchereria bancrofti (isolation & purification)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ADN des helminthes (génétique)</term><term>ADN des helminthes (sang)</term><term>Analyse de régression</term><term>Animaux</term><term>Données de séquences moléculaires</term><term>Filariose lymphatique (diagnostic)</term><term>Filariose lymphatique (sang)</term><term>Humains</term><term>Reproductibilité des résultats</term><term>Réaction de polymérisation en chaîne ()</term><term>Sensibilité et spécificité</term><term>Sondes d'ADN</term><term>Séquence nucléotidique</term><term>Séquences répétées d'acides nucléiques</term><term>Test ELISA ()</term><term>Type II site-specific deoxyribonuclease</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="genetics" xml:lang="en"><term>DNA, Helminth</term></keywords><keywords scheme="MESH" type="chemical" xml:lang="en"><term>DNA Probes</term><term>Deoxyribonucleases, Type II Site-Specific</term></keywords><keywords scheme="MESH" qualifier="blood" xml:lang="en"><term>Elephantiasis, Filarial</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>Wuchereria bancrofti</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>ADN des helminthes</term><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="methods" xml:lang="en"><term>Enzyme-Linked Immunosorbent Assay</term><term>Polymerase Chain Reaction</term></keywords><keywords scheme="MESH" qualifier="sang" xml:lang="fr"><term>ADN des helminthes</term><term>Filariose lymphatique</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Base Sequence</term><term>Humans</term><term>Molecular Sequence Data</term><term>Regression Analysis</term><term>Repetitive Sequences, Nucleic Acid</term><term>Reproducibility of Results</term><term>Sensitivity and Specificity</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Analyse de régression</term><term>Animaux</term><term>Données de séquences moléculaires</term><term>Humains</term><term>Reproductibilité des résultats</term><term>Réaction de polymérisation en chaîne</term><term>Sensibilité et spécificité</term><term>Sondes d'ADN</term><term>Séquence nucléotidique</term><term>Séquences répétées d'acides nucléiques</term><term>Test ELISA</term><term>Type II site-specific deoxyribonuclease</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">A quantitative, competitive polymerase chain reaction (QC-PCR) assay for the sensitive detection of Wuchereria bancrofti DNA was developed. A competitor sequence was constructed by an exchange of nucleotides in the Wuchereria-specific Ssp I repeat. The PCR products were hybridized to specific DNA probes and their amounts, determined by an enzyme-linked immunosorbent assay (ELISA). In laboratory-prepared samples the QC-PCR-ELISA assay was capable of detecting the amount of DNA equivalent to 0.1 microfilaria (mf) added to 200 microl of blood lysate. The assay was also tested on 78 blood samples collected in endemic areas in Egypt. All 28 samples that were positive both for mf and for circulating antigen were also QC-PCR-ELISA-positive. In addition, one mf-negative but antigen-positive sample was also positive as determined by QC-PCR-ELISA. A positive correlation of mf density with the QC-PCR-ELISA was observed. Samples containing 10 or fewer mf/ml had a mean relative amount of Ssp I PCR product of 19.7 units, whereas samples with 11-100 mf/ml had a mean of 36.3 units and those with more than 100 mf/ml had a mean of 84.6 units. Because of the high standard deviation within each group, estimates of worm burdens in infected individuals using the QC-PCR-ELISA are not recommended. However, we present data indicating that the W. bancrofti QC-PCR-ELISA is a powerful new tool for evaluation of parasitic loads for community-based diagnosis of bancroftian filariasis.</div></front></TEI><affiliations><list><country><li>États-Unis</li></country><region><li>Massachusetts</li></region></list><tree><noCountry><name sortKey="Kamal, I H" sort="Kamal, I H" uniqKey="Kamal I" first="I H" last="Kamal">I H Kamal</name><name sortKey="Liu, X" sort="Liu, X" uniqKey="Liu X" first="X" last="Liu">X. Liu</name><name sortKey="Lizotte Waniewski, M" sort="Lizotte Waniewski, M" uniqKey="Lizotte Waniewski M" first="M" last="Lizotte-Waniewski">M. Lizotte-Waniewski</name><name sortKey="Ramzy, R M" sort="Ramzy, R M" uniqKey="Ramzy R" first="R M" last="Ramzy">R M Ramzy</name><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="Fischer, P" sort="Fischer, P" uniqKey="Fischer P" first="P" last="Fischer">P. Fischer</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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