Validation of a multiplex reverse transcriptase PCR ELISA for the detection of 19 respiratory tract pathogens
Identifieur interne : 000179 ( France/Analysis ); précédent : 000178; suivant : 000180Validation of a multiplex reverse transcriptase PCR ELISA for the detection of 19 respiratory tract pathogens
Auteurs : W. Puppe [Allemagne] ; J. Weigl [Allemagne] ; B. Gröndahl [Allemagne] ; M. Knuf [Allemagne] ; S. Rockahr [Allemagne] ; P. Von Bismarck [Allemagne] ; G. Aron [Pays-Bas] ; H. G. M. Niesters [Pays-Bas] ; A. D. M. E. Osterhaus [Pays-Bas] ; H.-J. Schmitt [Allemagne, France]Source :
- Infection [ 0300-8126 ] ; 2013-02-01.
English descriptors
- KwdEn :
- Adenovirus (AV), Bordetella parapertussis ( Bpp ), Bordetella pertussis ( Bp ), Chlamydophila pneumoniae ( Cpn ), Coronavirus (CV), Diagnosis, Enterovirus (EV), Human metapneumovirus (MPV), Influenza virus type A (IVA), Influenza virus type B (IVB), Legionella pneumophila ( Lpn ), Mycoplasma pneumoniae ( Mpn ), Parainfluenza virus type 1 (PIV1), Parainfluenza virus type 2 (PIV2), Parainfluenza virus type 3 (PIV3), Parainfluenza virus type 4 (PIV4), Reovirus (ReoV), Respiratory syncytial virus (RSV), Respiratory tract infections, Rhinovirus (RV), Sensitivity, Validation, m-RT-PCR-ELISA.
Abstract
Abstract: Introduction: Since acute respiratory tract infections inflict a high burden of disease in children worldwide, a multiplex reverse transcription polymerase chain reaction combined with a microwell hybridization assay (m-RT-PCR–ELISA) to detect 19 different respiratory pathogens was developed and validated. Methods: A total of 430 respiratory specimens were retrospectively tested in parallel by both the advanced 19-valent m-RT-PCR–ELISA as well as by culture or individual RT-PCR assays used in clinical routine. Results: The mean (median) sensitivity of the m-RT-PCR–ELISA in the retrospective test was 93.3% (95.1%; range 83.3–100 %), and the mean (median) specificity was 99.8 and 100 % (range 98.6–100 %), respectively. The mean positive predictive value was 99.3 % (range 93.4–100 %) and the mean negative predictive value was 95.3 % (range 98.4–100 %). Feasibility and clinical value of the 19-valent method was prospectively shown on 16,231 incoming clinical specimens from patients between 0 and 16 years of age with acute respiratory tract infections admitted to pediatric hospitals or private practices from October 2003 to June 2010 in three regions in Germany (Kiel, Mainz, Freiburg; Freiburg to June 2007 only). At least one microorganism was detected in 10,765 of 16,231 (66.3 %) clinical specimens: 5,044 RV, 1,999 RSV, 1,286 AV, 944 EV, 737 seasonal IVA, 173 pandemic IVA H1N1-2009, 899 MPV, 518 CV, 383 PIV3, 268 PIV1, 259 Mpn, 205 IVB, 164 PIV2, 144 PIV4, 103 Bp, 29 Cpn and 29 Bpp, while reovirus and Lpn were not present in these specimens from a pediatric population. More than one organism could be detected in 13.4 % of the specimens. Conclusions: The m-RT-PCR–ELISA evaluated here improves the spectrum for diagnosing respiratory infections and is a feasible instrument for individual diagnostic and epidemiological studies.
Url:
DOI: 10.1007/s15010-012-0298-6
Affiliations:
- Allemagne, France, Pays-Bas
- Basse-Saxe, Groningue (province), Hollande-Méridionale, Schleswig-Holstein, Île-de-France
- Groningue, Hanovre, Kiel, Paris, Rotterdam
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ISTEX:2FC24E720BD3D59CDC5EC064D8A0FF1F98EA73C4Le document en format XML
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Niesters</name><affiliation wicri:level="3"><country xml:lang="fr">Pays-Bas</country><wicri:regionArea>Department of Virology, Erasmus MC., ’s Gravendijkwal 230, 3000 CA, Rotterdam</wicri:regionArea><placeName><settlement type="city">Rotterdam</settlement><region nuts="2" type="province">Hollande-Méridionale</region></placeName></affiliation><affiliation wicri:level="3"><country xml:lang="fr">Pays-Bas</country><wicri:regionArea>Department of Medical Microbiology/Virology, University Medical Center Groningen, Hanzeplein 1, 9713 GZ, Groningen</wicri:regionArea><placeName><settlement type="city">Groningue</settlement><region nuts="2" type="province">Groningue (province)</region></placeName></affiliation></author><author><name sortKey="Osterhaus, A D M E" sort="Osterhaus, A D M E" uniqKey="Osterhaus A" first="A. D. M. E." last="Osterhaus">A. D. M. E. Osterhaus</name><affiliation wicri:level="3"><country xml:lang="fr">Pays-Bas</country><wicri:regionArea>Department of Virology, Erasmus MC., ’s Gravendijkwal 230, 3000 CA, Rotterdam</wicri:regionArea><placeName><settlement type="city">Rotterdam</settlement><region nuts="2" type="province">Hollande-Méridionale</region></placeName></affiliation></author><author><name sortKey="Schmitt, H J" sort="Schmitt, H J" uniqKey="Schmitt H" first="H.-J." last="Schmitt">H.-J. Schmitt</name><affiliation wicri:level="1"><country xml:lang="fr">Allemagne</country><wicri:regionArea>Pediatric Infectious Diseases, Department of Pediatrics, University Medicine, Langenbeckstr. 1, 55101, Mainz</wicri:regionArea><wicri:noRegion>55101, Mainz</wicri:noRegion><wicri:noRegion>Mainz</wicri:noRegion></affiliation><affiliation wicri:level="3"><country xml:lang="fr">France</country><wicri:regionArea>Pfizer, 23–25 Avenue du Dr. Lannelongue, 75014, Paris</wicri:regionArea><placeName><region type="region" nuts="2">Île-de-France</region><settlement type="city">Paris</settlement></placeName></affiliation></author></analytic><monogr></monogr><series><title level="j">Infection</title><title level="j" type="sub">A Journal of Infectious Disease</title><title level="j" type="abbrev">Infection</title><idno type="ISSN">0300-8126</idno><idno type="eISSN">1439-0973</idno><imprint><publisher>Springer-Verlag</publisher><pubPlace>Berlin/Heidelberg</pubPlace><date type="published" when="2013-02-01">2013-02-01</date><biblScope unit="volume">41</biblScope><biblScope unit="issue">1</biblScope><biblScope unit="page" from="77">77</biblScope><biblScope unit="page" to="91">91</biblScope></imprint><idno type="ISSN">0300-8126</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0300-8126</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Adenovirus (AV)</term><term>Bordetella parapertussis ( Bpp )</term><term>Bordetella pertussis ( Bp )</term><term>Chlamydophila pneumoniae ( Cpn )</term><term>Coronavirus (CV)</term><term>Diagnosis</term><term>Enterovirus (EV)</term><term>Human metapneumovirus (MPV)</term><term>Influenza virus type A (IVA)</term><term>Influenza virus type B (IVB)</term><term>Legionella pneumophila ( Lpn )</term><term>Mycoplasma pneumoniae ( Mpn )</term><term>Parainfluenza virus type 1 (PIV1)</term><term>Parainfluenza virus type 2 (PIV2)</term><term>Parainfluenza virus type 3 (PIV3)</term><term>Parainfluenza virus type 4 (PIV4)</term><term>Reovirus (ReoV)</term><term>Respiratory syncytial virus (RSV)</term><term>Respiratory tract infections</term><term>Rhinovirus (RV)</term><term>Sensitivity</term><term>Validation</term><term>m-RT-PCR-ELISA</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: Introduction: Since acute respiratory tract infections inflict a high burden of disease in children worldwide, a multiplex reverse transcription polymerase chain reaction combined with a microwell hybridization assay (m-RT-PCR–ELISA) to detect 19 different respiratory pathogens was developed and validated. Methods: A total of 430 respiratory specimens were retrospectively tested in parallel by both the advanced 19-valent m-RT-PCR–ELISA as well as by culture or individual RT-PCR assays used in clinical routine. Results: The mean (median) sensitivity of the m-RT-PCR–ELISA in the retrospective test was 93.3% (95.1%; range 83.3–100 %), and the mean (median) specificity was 99.8 and 100 % (range 98.6–100 %), respectively. The mean positive predictive value was 99.3 % (range 93.4–100 %) and the mean negative predictive value was 95.3 % (range 98.4–100 %). Feasibility and clinical value of the 19-valent method was prospectively shown on 16,231 incoming clinical specimens from patients between 0 and 16 years of age with acute respiratory tract infections admitted to pediatric hospitals or private practices from October 2003 to June 2010 in three regions in Germany (Kiel, Mainz, Freiburg; Freiburg to June 2007 only). At least one microorganism was detected in 10,765 of 16,231 (66.3 %) clinical specimens: 5,044 RV, 1,999 RSV, 1,286 AV, 944 EV, 737 seasonal IVA, 173 pandemic IVA H1N1-2009, 899 MPV, 518 CV, 383 PIV3, 268 PIV1, 259 Mpn, 205 IVB, 164 PIV2, 144 PIV4, 103 Bp, 29 Cpn and 29 Bpp, while reovirus and Lpn were not present in these specimens from a pediatric population. More than one organism could be detected in 13.4 % of the specimens. Conclusions: The m-RT-PCR–ELISA evaluated here improves the spectrum for diagnosing respiratory infections and is a feasible instrument for individual diagnostic and epidemiological studies.</div></front></TEI><affiliations><list><country><li>Allemagne</li><li>France</li><li>Pays-Bas</li></country><region><li>Basse-Saxe</li><li>Groningue (province)</li><li>Hollande-Méridionale</li><li>Schleswig-Holstein</li><li>Île-de-France</li></region><settlement><li>Groningue</li><li>Hanovre</li><li>Kiel</li><li>Paris</li><li>Rotterdam</li></settlement></list><tree><country name="Allemagne"><region name="Schleswig-Holstein"><name sortKey="Puppe, W" sort="Puppe, W" uniqKey="Puppe W" first="W." last="Puppe">W. Puppe</name></region><name sortKey="Grondahl, B" sort="Grondahl, B" uniqKey="Grondahl B" first="B." last="Gröndahl">B. Gröndahl</name><name sortKey="Grondahl, B" sort="Grondahl, B" uniqKey="Grondahl B" first="B." last="Gröndahl">B. Gröndahl</name><name sortKey="Knuf, M" sort="Knuf, M" uniqKey="Knuf M" first="M." last="Knuf">M. Knuf</name><name sortKey="Puppe, W" sort="Puppe, W" uniqKey="Puppe W" first="W." last="Puppe">W. Puppe</name><name sortKey="Puppe, W" sort="Puppe, W" uniqKey="Puppe W" first="W." last="Puppe">W. Puppe</name><name sortKey="Rockahr, S" sort="Rockahr, S" uniqKey="Rockahr S" first="S." last="Rockahr">S. Rockahr</name><name sortKey="Schmitt, H J" sort="Schmitt, H J" uniqKey="Schmitt H" first="H.-J." last="Schmitt">H.-J. Schmitt</name><name sortKey="Von Bismarck, P" sort="Von Bismarck, P" uniqKey="Von Bismarck P" first="P." last="Von Bismarck">P. Von Bismarck</name><name sortKey="Weigl, J" sort="Weigl, J" uniqKey="Weigl J" first="J." last="Weigl">J. Weigl</name><name sortKey="Weigl, J" sort="Weigl, J" uniqKey="Weigl J" first="J." last="Weigl">J. Weigl</name></country><country name="Pays-Bas"><region name="Hollande-Méridionale"><name sortKey="Aron, G" sort="Aron, G" uniqKey="Aron G" first="G." last="Aron">G. Aron</name></region><name sortKey="Niesters, H G M" sort="Niesters, H G M" uniqKey="Niesters H" first="H. G. M." last="Niesters">H. G. M. Niesters</name><name sortKey="Niesters, H G M" sort="Niesters, H G M" uniqKey="Niesters H" first="H. G. M." last="Niesters">H. G. M. Niesters</name><name sortKey="Osterhaus, A D M E" sort="Osterhaus, A D M E" uniqKey="Osterhaus A" first="A. D. M. E." last="Osterhaus">A. D. M. E. Osterhaus</name></country><country name="France"><region name="Île-de-France"><name sortKey="Schmitt, H J" sort="Schmitt, H J" uniqKey="Schmitt H" first="H.-J." last="Schmitt">H.-J. Schmitt</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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