Analytical Comparison of In Vitro-Spiked Human Serum and Plasma for PCR-Based Detection of Aspergillus fumigatus DNA: a Study by the European Aspergillus PCR Initiative
Identifieur interne : 002726 ( Ncbi/Curation ); précédent : 002725; suivant : 002727Analytical Comparison of In Vitro-Spiked Human Serum and Plasma for PCR-Based Detection of Aspergillus fumigatus DNA: a Study by the European Aspergillus PCR Initiative
Auteurs : Juergen Loeffler [Allemagne] ; Carlo Mengoli [Italie] ; Jan Springer [Allemagne] ; Stéphane Bretagne [France] ; Manuel Cuenca-Estrella [Espagne] ; Lena Klingspor [Suède] ; Katrien Lagrou [Belgique] ; Willem J. G. Melchers [Pays-Bas] ; C. Oliver Morton [Australie] ; Rosemary A. Barnes [Royaume-Uni] ; J. Peter Donnelly [Pays-Bas] ; P. Lewis White [Royaume-Uni]Source :
- Journal of Clinical Microbiology [ 0095-1137 ] ; 2015.
Descripteurs français
- KwdFr :
- ADN fongique (sang), Aspergillose (diagnostic), Aspergillose (microbiologie), Aspergillus fumigatus (génétique), Aspergillus fumigatus (isolement et purification), Facteurs temps, Humains, Modèles biologiques, Plasma sanguin (microbiologie), Prélèvement biologique (), Réaction de polymérisation en chaîne (), Sensibilité et spécificité, Techniques de diagnostic moléculaire ().
- MESH :
- diagnostic : Aspergillose.
- génétique : Aspergillus fumigatus.
- isolement et purification : Aspergillus fumigatus.
- microbiologie : Aspergillose, Plasma sanguin.
- sang : ADN fongique.
- Facteurs temps, Humains, Modèles biologiques, Prélèvement biologique, Réaction de polymérisation en chaîne, Sensibilité et spécificité, Techniques de diagnostic moléculaire.
English descriptors
- KwdEn :
- Aspergillosis (diagnosis), Aspergillosis (microbiology), Aspergillus fumigatus (genetics), Aspergillus fumigatus (isolation & purification), DNA, Fungal (blood), Humans, Models, Biological, Molecular Diagnostic Techniques (methods), Plasma (microbiology), Polymerase Chain Reaction (methods), Sensitivity and Specificity, Specimen Handling (methods), Time Factors.
- MESH :
- chemical , blood : DNA, Fungal.
- diagnosis : Aspergillosis.
- genetics : Aspergillus fumigatus.
- isolation & purification : Aspergillus fumigatus.
- methods : Molecular Diagnostic Techniques, Polymerase Chain Reaction, Specimen Handling.
- microbiology : Aspergillosis, Plasma.
- Humans, Models, Biological, Sensitivity and Specificity, Time Factors.
Abstract
The use of serum or plasma for
Url:
DOI: 10.1128/JCM.00906-15
PubMed: 26085614
PubMed Central: 4540929
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PMC:4540929Le document en format XML
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-Spiked Human Serum and Plasma for PCR-Based Detection of <named-content content-type="genus-species">Aspergillus fumigatus</named-content>
DNA: a Study by the European <named-content content-type="genus-species">Aspergillus</named-content>
PCR Initiative</title>
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-Spiked Human Serum and Plasma for PCR-Based Detection of <named-content content-type="genus-species">Aspergillus fumigatus</named-content>
DNA: a Study by the European <named-content content-type="genus-species">Aspergillus</named-content>
PCR Initiative</title>
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<author><name sortKey="Cuenca Estrella, Manuel" sort="Cuenca Estrella, Manuel" uniqKey="Cuenca Estrella M" first="Manuel" last="Cuenca-Estrella">Manuel Cuenca-Estrella</name>
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<author><name sortKey="Lagrou, Katrien" sort="Lagrou, Katrien" uniqKey="Lagrou K" first="Katrien" last="Lagrou">Katrien Lagrou</name>
<affiliation wicri:level="1"><nlm:aff id="aff6">National Reference Center for Mycosis, University Hospitals Leuven, Leuven, Belgium</nlm:aff>
<country xml:lang="fr">Belgique</country>
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<affiliation wicri:level="1"><nlm:aff id="aff7">Department of Microbiology and Immunology, KU Leuven, Leuven, Belgium</nlm:aff>
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<author><name sortKey="Melchers, Willem J G" sort="Melchers, Willem J G" uniqKey="Melchers W" first="Willem J. G." last="Melchers">Willem J. G. Melchers</name>
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<country xml:lang="fr">Pays-Bas</country>
<wicri:regionArea>Radboud University Medical Centre, Nijmegen</wicri:regionArea>
<placeName><settlement type="city">Nimègue</settlement>
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<author><name sortKey="Morton, C Oliver" sort="Morton, C Oliver" uniqKey="Morton C" first="C. Oliver" last="Morton">C. Oliver Morton</name>
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<author><name sortKey="Barnes, Rosemary A" sort="Barnes, Rosemary A" uniqKey="Barnes R" first="Rosemary A." last="Barnes">Rosemary A. Barnes</name>
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<author><name sortKey="Donnelly, J Peter" sort="Donnelly, J Peter" uniqKey="Donnelly J" first="J. Peter" last="Donnelly">J. Peter Donnelly</name>
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<author><name sortKey="White, P Lewis" sort="White, P Lewis" uniqKey="White P" first="P. Lewis" last="White">P. Lewis White</name>
<affiliation wicri:level="1"><nlm:aff id="aff11">Public Health Wales Microbiology Cardiff, Cardiff, United Kingdom</nlm:aff>
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<series><title level="j">Journal of Clinical Microbiology</title>
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<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Aspergillosis (diagnosis)</term>
<term>Aspergillosis (microbiology)</term>
<term>Aspergillus fumigatus (genetics)</term>
<term>Aspergillus fumigatus (isolation & purification)</term>
<term>DNA, Fungal (blood)</term>
<term>Humans</term>
<term>Models, Biological</term>
<term>Molecular Diagnostic Techniques (methods)</term>
<term>Plasma (microbiology)</term>
<term>Polymerase Chain Reaction (methods)</term>
<term>Sensitivity and Specificity</term>
<term>Specimen Handling (methods)</term>
<term>Time Factors</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>ADN fongique (sang)</term>
<term>Aspergillose (diagnostic)</term>
<term>Aspergillose (microbiologie)</term>
<term>Aspergillus fumigatus (génétique)</term>
<term>Aspergillus fumigatus (isolement et purification)</term>
<term>Facteurs temps</term>
<term>Humains</term>
<term>Modèles biologiques</term>
<term>Plasma sanguin (microbiologie)</term>
<term>Prélèvement biologique ()</term>
<term>Réaction de polymérisation en chaîne ()</term>
<term>Sensibilité et spécificité</term>
<term>Techniques de diagnostic moléculaire ()</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="blood" xml:lang="en"><term>DNA, Fungal</term>
</keywords>
<keywords scheme="MESH" qualifier="diagnosis" xml:lang="en"><term>Aspergillosis</term>
</keywords>
<keywords scheme="MESH" qualifier="diagnostic" xml:lang="fr"><term>Aspergillose</term>
</keywords>
<keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Aspergillus fumigatus</term>
</keywords>
<keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Aspergillus fumigatus</term>
</keywords>
<keywords scheme="MESH" qualifier="isolation & purification" xml:lang="en"><term>Aspergillus fumigatus</term>
</keywords>
<keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Aspergillus fumigatus</term>
</keywords>
<keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Molecular Diagnostic Techniques</term>
<term>Polymerase Chain Reaction</term>
<term>Specimen Handling</term>
</keywords>
<keywords scheme="MESH" qualifier="microbiologie" xml:lang="fr"><term>Aspergillose</term>
<term>Plasma sanguin</term>
</keywords>
<keywords scheme="MESH" qualifier="microbiology" xml:lang="en"><term>Aspergillosis</term>
<term>Plasma</term>
</keywords>
<keywords scheme="MESH" qualifier="sang" xml:lang="fr"><term>ADN fongique</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Humans</term>
<term>Models, Biological</term>
<term>Sensitivity and Specificity</term>
<term>Time Factors</term>
</keywords>
<keywords scheme="MESH" xml:lang="fr"><term>Facteurs temps</term>
<term>Humains</term>
<term>Modèles biologiques</term>
<term>Prélèvement biologique</term>
<term>Réaction de polymérisation en chaîne</term>
<term>Sensibilité et spécificité</term>
<term>Techniques de diagnostic moléculaire</term>
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<front><div type="abstract" xml:lang="en"><p>The use of serum or plasma for <named-content content-type="genus-species">Aspergillus</named-content>
PCR testing facilitates automated and standardized technology. Recommendations for serum testing are available, and while serum and plasma are regularly considered interchangeable for use in fungal diagnostics, differences in galactomannan enzyme immunoassay (GM-EIA) performance have been reported and are attributed to clot formation. Therefore, it is important to assess plasma PCR testing to determine if previous recommendations for serum are applicable and also to compare analytical performance with that of serum PCR. Molecular methods testing serum and plasma were compared through multicenter distribution of quality control panels, with additional studies to investigate the effect of clot formation and blood fractionation on DNA availability. Analytical sensitivity and time to positivity (TTP) were compared, and a regression analysis was performed to identify variables that enhanced plasma PCR performance. When testing plasma, sample volume, preextraction-to-postextraction volume ratio, PCR volume, duplicate testing, and the use of an internal control for PCR were positively associated with performance. When whole-blood samples were spiked and then fractionated, the analytical sensitivity and TTP were superior when testing plasma. Centrifugation had no effect on DNA availability, whereas the presence of clot material significantly lowered the concentration (<italic>P</italic>
= 0.028). Technically, there are no major differences in the molecular processing of serum and plasma, but the formation of clot material potentially reduces available DNA in serum. During disease, <named-content content-type="genus-species">Aspergillus</named-content>
DNA burdens in blood are often at the limits of PCR performance. Using plasma might improve performance while maintaining the methodological simplicity of serum testing.</p>
</div>
</front>
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