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Published in: Archives of Virology 4/2014

01-04-2014 | Original Article

Detection of respiratory viruses using a multiplex real-time PCR assay in Germany, 2009/10

Authors: Sibylle Bierbaum, Johannes Forster, Reinhard Berner, Gerta Rücker, Gernot Rohde, Dieter Neumann-Haefelin, Marcus Panning, CAPNETZ study group

Published in: Archives of Virology | Issue 4/2014

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Abstract

The aim of this study was to determine the prevalence of respiratory viruses and to prospectively evaluate the performance of the fast-track diagnostics (FTD) respiratory pathogens multiplex PCR assay shortly after the 2009/10 influenza pandemic. Highly sensitive monoplex real-time PCR assays served as references. Discrepant results were further analyzed by the xTAG RVP Fast assay. A total of 369 respiratory samples from children and adults were collected prospectively in Germany from December 2009 until June 2010. The sensitivity and specificity of the FTD assay after resolution of discrepant results was 92.2 % and 99.5 %, respectively. Lowest specificity of the FTD assay was observed for human bocavirus. Multiple detections were recorded in 33/369 (8.9 %) of the samples by monoplex PCR and in 43/369 (11.7 %) using the FTD assay. The most prevalent viruses were respiratory syncytial virus and human metapneumovirus. Only pandemic influenza virus A/H1N1 (2009), and not seasonal influenza virus, was detected. Viruses other than influenza virus accounted for the majority of acute respiratory infections. The FTD assay can be easily implemented in general diagnostic laboratories and facilitate the optimization of patient-management schemes.
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Literature
1.
go back to reference Anderson TP, Werno AM, Barratt K, Mahagamasekera P, Murdoch DR, Jennings LC (2013) Comparison of four multiplex PCR assays for the detection of viral pathogens in respiratory specimens. J Virol Methods 191:118–121PubMedCrossRef Anderson TP, Werno AM, Barratt K, Mahagamasekera P, Murdoch DR, Jennings LC (2013) Comparison of four multiplex PCR assays for the detection of viral pathogens in respiratory specimens. J Virol Methods 191:118–121PubMedCrossRef
2.
go back to reference Bierbaum S, Konigsfeld N, Besazza N, Blessing K, Rucker G, Kontny U, Berner R, Schumacher M, Forster J, Falcone V, Van De Sand C, Essig A, Huzly D, Rohde G, Neumann-Haefelin D, Panning M (2012) Performance of a novel microarray multiplex PCR for the detection of 23 respiratory pathogens (SYMP-ARI study). Eur J Clin Microbiol Infect Dis 31:2851–2861PubMedCrossRef Bierbaum S, Konigsfeld N, Besazza N, Blessing K, Rucker G, Kontny U, Berner R, Schumacher M, Forster J, Falcone V, Van De Sand C, Essig A, Huzly D, Rohde G, Neumann-Haefelin D, Panning M (2012) Performance of a novel microarray multiplex PCR for the detection of 23 respiratory pathogens (SYMP-ARI study). Eur J Clin Microbiol Infect Dis 31:2851–2861PubMedCrossRef
3.
go back to reference Bonzel L, Tenenbaum T, Schroten H, Schildgen O, Schweitzer-Krantz S, Adams O (2008) Frequent detection of viral coinfection in children hospitalized with acute respiratory tract infection using a real-time polymerase chain reaction. Pediatr Infect Dis J 27:589–594PubMedCrossRef Bonzel L, Tenenbaum T, Schroten H, Schildgen O, Schweitzer-Krantz S, Adams O (2008) Frequent detection of viral coinfection in children hospitalized with acute respiratory tract infection using a real-time polymerase chain reaction. Pediatr Infect Dis J 27:589–594PubMedCrossRef
4.
go back to reference Brittain-Long R, Westin J, Olofsson S, Lindh M, Andersson LM (2011) Access to a polymerase chain reaction assay method targeting 13 respiratory viruses can reduce antibiotics: a randomised, controlled trial. BMC Med 9:44PubMedCentralPubMedCrossRef Brittain-Long R, Westin J, Olofsson S, Lindh M, Andersson LM (2011) Access to a polymerase chain reaction assay method targeting 13 respiratory viruses can reduce antibiotics: a randomised, controlled trial. BMC Med 9:44PubMedCentralPubMedCrossRef
5.
go back to reference Brunstein JD, Cline CL, Mckinney S, Thomas E (2008) Evidence from multiplex molecular assays for complex multipathogen interactions in acute respiratory infections. J Clin Microbiol 46:97–102PubMedCentralPubMedCrossRef Brunstein JD, Cline CL, Mckinney S, Thomas E (2008) Evidence from multiplex molecular assays for complex multipathogen interactions in acute respiratory infections. J Clin Microbiol 46:97–102PubMedCentralPubMedCrossRef
6.
go back to reference Dreier J, Stormer M, Kleesiek K (2005) Use of bacteriophage MS2 as an internal control in viral reverse transcription-PCR assays. J Clin Microbiol 43:4551–4557PubMedCentralPubMedCrossRef Dreier J, Stormer M, Kleesiek K (2005) Use of bacteriophage MS2 as an internal control in viral reverse transcription-PCR assays. J Clin Microbiol 43:4551–4557PubMedCentralPubMedCrossRef
8.
go back to reference Druce J, Garcia K, Tran T, Papadakis G, Birch C (2012) Evaluation of swabs, transport media, and specimen transport conditions for optimal detection of viruses by PCR. J Clin Microbiol 50:1064–1065PubMedCentralPubMedCrossRef Druce J, Garcia K, Tran T, Papadakis G, Birch C (2012) Evaluation of swabs, transport media, and specimen transport conditions for optimal detection of viruses by PCR. J Clin Microbiol 50:1064–1065PubMedCentralPubMedCrossRef
9.
go back to reference Faux CE, Arden KE, Lambert SB, Nissen MD, Nolan TM, Chang AB, Sloots TP, Mackay IM (2011) Usefulness of published PCR primers in detecting human rhinovirus infection. Emerg Infect Dis 17:296–298PubMedCentralPubMedCrossRef Faux CE, Arden KE, Lambert SB, Nissen MD, Nolan TM, Chang AB, Sloots TP, Mackay IM (2011) Usefulness of published PCR primers in detecting human rhinovirus infection. Emerg Infect Dis 17:296–298PubMedCentralPubMedCrossRef
10.
go back to reference Fleiss JL, Cohen J, Everitt BS (1969) Large sample standard errors of kappa and weighted kappa. Psychol Bull 72:323–327CrossRef Fleiss JL, Cohen J, Everitt BS (1969) Large sample standard errors of kappa and weighted kappa. Psychol Bull 72:323–327CrossRef
11.
go back to reference Gadsby NJ, Hardie A, Claas EC, Templeton KE (2010) Comparison of the luminex respiratory virus panel fast assay with in-house real-time PCR for respiratory viral infection diagnosis. J Clin Microbiol 48:2213–2216PubMedCentralPubMedCrossRef Gadsby NJ, Hardie A, Claas EC, Templeton KE (2010) Comparison of the luminex respiratory virus panel fast assay with in-house real-time PCR for respiratory viral infection diagnosis. J Clin Microbiol 48:2213–2216PubMedCentralPubMedCrossRef
12.
go back to reference Gharabaghi F, Hawan A, Drews SJ, Richardson SE (2011) Evaluation of multiple commercial molecular and conventional diagnostic assays for the detection of respiratory viruses in children. Clin Microbiol Infect 17:1900–1906PubMedCrossRef Gharabaghi F, Hawan A, Drews SJ, Richardson SE (2011) Evaluation of multiple commercial molecular and conventional diagnostic assays for the detection of respiratory viruses in children. Clin Microbiol Infect 17:1900–1906PubMedCrossRef
13.
go back to reference Gunson RN, Collins TC, Carman WF (2005) Real-time RT-PCR detection of 12 respiratory viral infections in four triplex reactions. J Clin Virol 33:341–344PubMedCrossRef Gunson RN, Collins TC, Carman WF (2005) Real-time RT-PCR detection of 12 respiratory viral infections in four triplex reactions. J Clin Virol 33:341–344PubMedCrossRef
14.
go back to reference Hoffmann J, Rabezanahary H, Randriamarotia M, Ratsimbasoa A, Najjar J, Vernet G, Contamin B, Paranhos-Baccala G (2012) Viral and atypical bacterial etiology of acute respiratory infections in children under 5 years old living in a rural tropical area of Madagascar. PLoS One 7:e43666PubMedCentralPubMedCrossRef Hoffmann J, Rabezanahary H, Randriamarotia M, Ratsimbasoa A, Najjar J, Vernet G, Contamin B, Paranhos-Baccala G (2012) Viral and atypical bacterial etiology of acute respiratory infections in children under 5 years old living in a rural tropical area of Madagascar. PLoS One 7:e43666PubMedCentralPubMedCrossRef
15.
16.
go back to reference Kim JK, Jeon JS, Kim JW, Rheem I (2013) Epidemiology of respiratory viral infection using multiplex rt-PCR in Cheonan, Korea (2006-2010). J Microbiol Biotechnol 23:267–273PubMedCrossRef Kim JK, Jeon JS, Kim JW, Rheem I (2013) Epidemiology of respiratory viral infection using multiplex rt-PCR in Cheonan, Korea (2006-2010). J Microbiol Biotechnol 23:267–273PubMedCrossRef
17.
go back to reference Dm Lamson, Griesemer S, Fuschino M, St George K (2012) Phylogenetic analysis of human metapneumovirus from New York State patients during February through April 2010. J Clin Virol 53:256–258CrossRef Dm Lamson, Griesemer S, Fuschino M, St George K (2012) Phylogenetic analysis of human metapneumovirus from New York State patients during February through April 2010. J Clin Virol 53:256–258CrossRef
18.
go back to reference Li J, Mao NY, Zhang C, Yang MJ, Wang M, Xu WB, Ma XJ (2012) The development of a GeXP-based multiplex reverse transcription-PCR assay for simultaneous detection of sixteen human respiratory virus types/subtypes. BMC Infect Dis 12:189PubMedCentralPubMedCrossRef Li J, Mao NY, Zhang C, Yang MJ, Wang M, Xu WB, Ma XJ (2012) The development of a GeXP-based multiplex reverse transcription-PCR assay for simultaneous detection of sixteen human respiratory virus types/subtypes. BMC Infect Dis 12:189PubMedCentralPubMedCrossRef
19.
go back to reference Panning M, Baumgarte S, Laue T, Bierbaum S, Raith S, Drexler JF, Helmer A, Falcone-Kapper V, Kochs G, Campe H, Huzly D, Eis-Hubinger AM, Drosten C (2011) Singleplex real-time RT-PCR for detection of influenza A virus and simultaneous differentiation of A/H1N1v and evaluation of the RealStar influenza kit. J Clin Virol 50:171–174PubMedCrossRef Panning M, Baumgarte S, Laue T, Bierbaum S, Raith S, Drexler JF, Helmer A, Falcone-Kapper V, Kochs G, Campe H, Huzly D, Eis-Hubinger AM, Drosten C (2011) Singleplex real-time RT-PCR for detection of influenza A virus and simultaneous differentiation of A/H1N1v and evaluation of the RealStar influenza kit. J Clin Virol 50:171–174PubMedCrossRef
20.
go back to reference Panning M, Charrel RN, Donoso Mantke O, Landt O, Niedrig M, Drosten C (2009) Coordinated implementation of chikungunya virus reverse transcription-PCR. Emerg Infect Dis 15:469–471PubMedCentralPubMedCrossRef Panning M, Charrel RN, Donoso Mantke O, Landt O, Niedrig M, Drosten C (2009) Coordinated implementation of chikungunya virus reverse transcription-PCR. Emerg Infect Dis 15:469–471PubMedCentralPubMedCrossRef
21.
go back to reference Pretorius MA, Madhi SA, Cohen C, Naidoo D, Groome M, Moyes J, Buys A, Walaza S, Dawood H, Chhagan M, Haffjee S, Kahn K, Puren A, Venter M (2013) Respiratory viral coinfections identified by a 10-plex real-time reverse-transcription polymerase chain reaction assay in patients hospitalized with severe acute respiratory illness–South Africa, 2009-2010. J Infect Dis 206(Suppl 1):S159–S165CrossRef Pretorius MA, Madhi SA, Cohen C, Naidoo D, Groome M, Moyes J, Buys A, Walaza S, Dawood H, Chhagan M, Haffjee S, Kahn K, Puren A, Venter M (2013) Respiratory viral coinfections identified by a 10-plex real-time reverse-transcription polymerase chain reaction assay in patients hospitalized with severe acute respiratory illness–South Africa, 2009-2010. J Infect Dis 206(Suppl 1):S159–S165CrossRef
22.
go back to reference Sakthivel SK, Whitaker B, Lu X, Oliveira DB, Stockman LJ, Kamili S, Oberste MS, Erdman DD (2012) Comparison of fast-track diagnostics respiratory pathogens multiplex real-time RT-PCR assay with in-house singleplex assays for comprehensive detection of human respiratory viruses. J Virol Methods 185:259–266PubMedCrossRef Sakthivel SK, Whitaker B, Lu X, Oliveira DB, Stockman LJ, Kamili S, Oberste MS, Erdman DD (2012) Comparison of fast-track diagnostics respiratory pathogens multiplex real-time RT-PCR assay with in-house singleplex assays for comprehensive detection of human respiratory viruses. J Virol Methods 185:259–266PubMedCrossRef
23.
go back to reference Wernike K, Bonilauri P, Dauber M, Errington J, Leblanc N, Revilla-Fernandez S, Hjulsager C, Isaksson M, Stadejek T, Beer M, Hoffmann B (2012) Porcine reproductive and respiratory syndrome virus: interlaboratory ring trial to evaluate real-time reverse transcription polymerase chain reaction detection methods. J Vet Diagn Invest 24:855–866PubMedCrossRef Wernike K, Bonilauri P, Dauber M, Errington J, Leblanc N, Revilla-Fernandez S, Hjulsager C, Isaksson M, Stadejek T, Beer M, Hoffmann B (2012) Porcine reproductive and respiratory syndrome virus: interlaboratory ring trial to evaluate real-time reverse transcription polymerase chain reaction detection methods. J Vet Diagn Invest 24:855–866PubMedCrossRef
24.
go back to reference Yang Y, Wang Z, Ren L, Wang W, Vernet G, Paranhos-Baccala G, Jin Q, Wang J (2012) Influenza A/H1N1 2009 pandemic and respiratory virus infections, Beijing, 2009-2010. PLoS One 7:e45807PubMedCentralPubMedCrossRef Yang Y, Wang Z, Ren L, Wang W, Vernet G, Paranhos-Baccala G, Jin Q, Wang J (2012) Influenza A/H1N1 2009 pandemic and respiratory virus infections, Beijing, 2009-2010. PLoS One 7:e45807PubMedCentralPubMedCrossRef
Metadata
Title
Detection of respiratory viruses using a multiplex real-time PCR assay in Germany, 2009/10
Authors
Sibylle Bierbaum
Johannes Forster
Reinhard Berner
Gerta Rücker
Gernot Rohde
Dieter Neumann-Haefelin
Marcus Panning
CAPNETZ study group
Publication date
01-04-2014
Publisher
Springer Vienna
Published in
Archives of Virology / Issue 4/2014
Print ISSN: 0304-8608
Electronic ISSN: 1432-8798
DOI
https://doi.org/10.1007/s00705-013-1876-3

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