Migrating a lab-developed MERS-CoV real-time PCR to 3 “Sample to Result” systems: experiences on optimization and validation
Identifieur interne : 000460 ( Main/Exploration ); précédent : 000459; suivant : 000461Migrating a lab-developed MERS-CoV real-time PCR to 3 “Sample to Result” systems: experiences on optimization and validation
Auteurs : Glynis Frans ; Kurt Beuselinck ; Bart Peeters ; Marc Van Ranst ; Veroniek Saegeman ; Stefanie Desmet ; Katrien LagrouSource :
- Diagnostic Microbiology and Infectious Disease [ 0732-8893 ] ; 2019.
Descripteurs français
- KwdFr :
- ARN viral (génétique), Amorces ADN, Coronavirus du syndrome respiratoire du Moyen-Orient (génétique), Coronavirus du syndrome respiratoire du Moyen-Orient (isolement et purification), Infections à coronavirus (diagnostic), Réaction de polymérisation en chaine en temps réel (), Réaction de polymérisation en chaine en temps réel (normes), Techniques de laboratoire clinique.
- MESH :
- diagnostic : Infections à coronavirus.
- génétique : ARN viral, Coronavirus du syndrome respiratoire du Moyen-Orient.
- isolement et purification : Coronavirus du syndrome respiratoire du Moyen-Orient.
- normes : Réaction de polymérisation en chaine en temps réel.
- Amorces ADN, Réaction de polymérisation en chaine en temps réel, Techniques de laboratoire clinique.
English descriptors
- KwdEn :
- Clinical Laboratory Techniques, Coronavirus Infections (diagnosis), DNA Primers, Middle East Respiratory Syndrome Coronavirus (genetics), Middle East Respiratory Syndrome Coronavirus (isolation & purification), RNA, Viral (genetics), Real-Time Polymerase Chain Reaction (methods), Real-Time Polymerase Chain Reaction (standards).
- MESH :
- chemical , genetics : RNA, Viral.
- chemical : DNA Primers.
- diagnosis : Coronavirus Infections.
- genetics : Middle East Respiratory Syndrome Coronavirus.
- isolation & purification : Middle East Respiratory Syndrome Coronavirus.
- methods : Real-Time Polymerase Chain Reaction.
- standards : Real-Time Polymerase Chain Reaction.
- Clinical Laboratory Techniques.
Abstract
The goal of the study was to adapt our Middle East respiratory syndrome coronavirus (MERS-CoV) lab-developed test (LDT) to 3 “Sample to Result” (S2R) systems: BD MAX (BD), ELITe InGenius (ELITechGroup), and ARIES (Luminex).
The BD MAX and InGenius system allowed use of lab-developed primers and TaqMan probes, while ARIES required conversion to MultiCode primers for melting curve analysis. Each device required ≤1 day of training and assay optimization. No discordant results were noted after analysis of 32 External Quality Control (EQC) samples. On a 10-fold dilution series of a MERS-CoV–positive EQC sample, InGenius obtained the highest detection rate. Laboratory technicians rated the ARIES as the user-friendliest. It also required the least hands-on time. BD MAX had the lowest turnaround time and highest throughput.
While each device had distinguishing system properties with associated (dis)advantages, the 3 S2R systems were comparable in terms of assay development and validation.
Url:
DOI: 10.1016/j.diagmicrobio.2019.02.006
PubMed: 30929995
PubMed Central: 7127711
Affiliations:
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“Sample to Result” systems: experiences on optimization and validation</title><author><name sortKey="Frans, Glynis" sort="Frans, Glynis" uniqKey="Frans G" first="Glynis" last="Frans">Glynis Frans</name></author><author><name sortKey="Beuselinck, Kurt" sort="Beuselinck, Kurt" uniqKey="Beuselinck K" first="Kurt" last="Beuselinck">Kurt Beuselinck</name></author><author><name sortKey="Peeters, Bart" sort="Peeters, Bart" uniqKey="Peeters B" first="Bart" last="Peeters">Bart Peeters</name></author><author><name sortKey="Van Ranst, Marc" sort="Van Ranst, Marc" uniqKey="Van Ranst M" first="Marc" last="Van Ranst">Marc Van Ranst</name></author><author><name sortKey="Saegeman, Veroniek" sort="Saegeman, Veroniek" uniqKey="Saegeman V" first="Veroniek" last="Saegeman">Veroniek Saegeman</name></author><author><name sortKey="Desmet, Stefanie" sort="Desmet, Stefanie" uniqKey="Desmet S" first="Stefanie" last="Desmet">Stefanie Desmet</name></author><author><name sortKey="Lagrou, Katrien" sort="Lagrou, Katrien" uniqKey="Lagrou K" first="Katrien" last="Lagrou">Katrien Lagrou</name></author></analytic><series><title level="j">Diagnostic Microbiology and Infectious Disease</title><idno type="ISSN">0732-8893</idno><idno type="eISSN">1879-0070</idno><imprint><date when="2019">2019</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Clinical Laboratory Techniques</term><term>Coronavirus Infections (diagnosis)</term><term>DNA Primers</term><term>Middle East Respiratory Syndrome Coronavirus (genetics)</term><term>Middle East Respiratory Syndrome Coronavirus (isolation & purification)</term><term>RNA, Viral (genetics)</term><term>Real-Time Polymerase Chain Reaction (methods)</term><term>Real-Time Polymerase Chain Reaction (standards)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ARN viral (génétique)</term><term>Amorces ADN</term><term>Coronavirus du syndrome respiratoire du Moyen-Orient (génétique)</term><term>Coronavirus du syndrome respiratoire du Moyen-Orient (isolement et purification)</term><term>Infections à coronavirus (diagnostic)</term><term>Réaction de polymérisation en chaine en temps réel ()</term><term>Réaction de polymérisation en chaine en temps réel (normes)</term><term>Techniques de laboratoire clinique</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>RNA, Viral</term></keywords><keywords scheme="MESH" type="chemical" xml:lang="en"><term>DNA Primers</term></keywords><keywords scheme="MESH" qualifier="diagnosis" xml:lang="en"><term>Coronavirus Infections</term></keywords><keywords scheme="MESH" qualifier="diagnostic" xml:lang="fr"><term>Infections à coronavirus</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Middle East Respiratory Syndrome Coronavirus</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>ARN viral</term><term>Coronavirus du syndrome respiratoire du Moyen-Orient</term></keywords><keywords scheme="MESH" qualifier="isolation & purification" xml:lang="en"><term>Middle East Respiratory Syndrome Coronavirus</term></keywords><keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Coronavirus du syndrome respiratoire du Moyen-Orient</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Real-Time Polymerase Chain Reaction</term></keywords><keywords scheme="MESH" qualifier="normes" xml:lang="fr"><term>Réaction de polymérisation en chaine en temps réel</term></keywords><keywords scheme="MESH" qualifier="standards" xml:lang="en"><term>Real-Time Polymerase Chain Reaction</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Clinical Laboratory Techniques</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Amorces ADN</term><term>Réaction de polymérisation en chaine en temps réel</term><term>Techniques de laboratoire clinique</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>The goal of the study was to adapt our Middle East respiratory syndrome coronavirus (MERS-CoV) lab-developed test (LDT) to 3 “Sample to Result” (S2R) systems: BD MAX (BD), ELITe InGenius (ELITechGroup), and ARIES (Luminex).</p><p>The BD MAX and InGenius system allowed use of lab-developed primers and TaqMan probes, while ARIES required conversion to MultiCode primers for melting curve analysis. Each device required ≤1 day of training and assay optimization. No discordant results were noted after analysis of 32 External Quality Control (EQC) samples. On a 10-fold dilution series of a MERS-CoV–positive EQC sample, InGenius obtained the highest detection rate. Laboratory technicians rated the ARIES as the user-friendliest. It also required the least hands-on time. BD MAX had the lowest turnaround time and highest throughput.</p><p>While each device had distinguishing system properties with associated (dis)advantages, the 3 S2R systems were comparable in terms of assay development and validation.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Azhar, E I" uniqKey="Azhar E">E.I. Azhar</name></author><author><name sortKey="El Kafrawy, S A" uniqKey="El Kafrawy S">S.A. El-Kafrawy</name></author><author><name sortKey="Farraj, S A" uniqKey="Farraj S">S.A. 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Zaki</name></author><author><name sortKey="Van Boheemen, S" uniqKey="Van Boheemen S">S. van Boheemen</name></author><author><name sortKey="Bestebroer, T M" uniqKey="Bestebroer T">T.M. Bestebroer</name></author><author><name sortKey="Osterhaus, A D" uniqKey="Osterhaus A">A.D. Osterhaus</name></author><author><name sortKey="Fouchier, R A" uniqKey="Fouchier R">R.A. Fouchier</name></author></analytic></biblStruct></listBibl></div1></back></TEI><affiliations><list></list><tree><noCountry><name sortKey="Beuselinck, Kurt" sort="Beuselinck, Kurt" uniqKey="Beuselinck K" first="Kurt" last="Beuselinck">Kurt Beuselinck</name><name sortKey="Desmet, Stefanie" sort="Desmet, Stefanie" uniqKey="Desmet S" first="Stefanie" last="Desmet">Stefanie Desmet</name><name sortKey="Frans, Glynis" sort="Frans, Glynis" uniqKey="Frans G" first="Glynis" last="Frans">Glynis Frans</name><name sortKey="Lagrou, Katrien" sort="Lagrou, Katrien" uniqKey="Lagrou K" first="Katrien" last="Lagrou">Katrien Lagrou</name><name sortKey="Peeters, Bart" sort="Peeters, Bart" uniqKey="Peeters B" first="Bart" last="Peeters">Bart Peeters</name><name sortKey="Saegeman, Veroniek" sort="Saegeman, Veroniek" uniqKey="Saegeman V" first="Veroniek" last="Saegeman">Veroniek Saegeman</name><name sortKey="Van Ranst, Marc" sort="Van Ranst, Marc" uniqKey="Van Ranst M" first="Marc" last="Van Ranst">Marc Van Ranst</name></noCountry></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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