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Published in: Infection 1/2013

01-02-2013 | Clinical and Epidemiological Study

Validation of a multiplex reverse transcriptase PCR ELISA for the detection of 19 respiratory tract pathogens

Authors: W. Puppe, J. Weigl, B. Gröndahl, M. Knuf, S. Rockahr, P. von Bismarck, G. Aron, H. G. M. Niesters, A. D. M. E. Osterhaus, H.-J. Schmitt

Published in: Infection | Issue 1/2013

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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.
Literature
1.
go back to reference Leowski J. Mortality from acute respiratory infections in children under 5 years of age: global estimates. World Health Stat Q. 1986;39:138–44.PubMed Leowski J. Mortality from acute respiratory infections in children under 5 years of age: global estimates. World Health Stat Q. 1986;39:138–44.PubMed
2.
go back to reference Gröndahl B, Puppe W, Hoppe A, Kühne I, Weigl JA, Schmitt HJ. Rapid identification of nine microorganisms causing acute respiratory tract infections by single-tube multiplex reverse transcription-PCR: feasibility study. J Clin Microbiol. 1999;37:1–7.PubMed Gröndahl B, Puppe W, Hoppe A, Kühne I, Weigl JA, Schmitt HJ. Rapid identification of nine microorganisms causing acute respiratory tract infections by single-tube multiplex reverse transcription-PCR: feasibility study. J Clin Microbiol. 1999;37:1–7.PubMed
3.
go back to reference Puppe W, Weigl JA, Aron G, Gröndahl B, Schmitt HJ, Niesters HG, Groen J. Evaluation of a multiplex reverse transcriptase PCR ELISA for the detection of nine respiratory tract pathogens. J Clin Virol. 2004;30:165–74.PubMedCrossRef Puppe W, Weigl JA, Aron G, Gröndahl B, Schmitt HJ, Niesters HG, Groen J. Evaluation of a multiplex reverse transcriptase PCR ELISA for the detection of nine respiratory tract pathogens. J Clin Virol. 2004;30:165–74.PubMedCrossRef
4.
go back to reference Brunstein JD, Cline CL, McKinney S, Thomas E. Evidence from multiplex molecular assays for complex multipathogen interactions in acute respiratory infections. J Clin Microbiol. 2008;46:97–102.PubMedCrossRef Brunstein JD, Cline CL, McKinney S, Thomas E. Evidence from multiplex molecular assays for complex multipathogen interactions in acute respiratory infections. J Clin Microbiol. 2008;46:97–102.PubMedCrossRef
5.
go back to reference Gruteke P, Glas AS, Dierdorp M, Vreede WB, Pilon JW, Bruisten SM. Practical implementation of a multiplex PCR for acute respiratory tract infections in children. J Clin Microbiol. 2004;42:5596–603.PubMedCrossRef Gruteke P, Glas AS, Dierdorp M, Vreede WB, Pilon JW, Bruisten SM. Practical implementation of a multiplex PCR for acute respiratory tract infections in children. J Clin Microbiol. 2004;42:5596–603.PubMedCrossRef
6.
go back to reference Kehl SC, Henrickson KJ, Hua W, Fan J. Evaluation of the Hexaplex assay for detection of respiratory viruses in children. J Clin Microbiol. 2001;39:1696–701.PubMedCrossRef Kehl SC, Henrickson KJ, Hua W, Fan J. Evaluation of the Hexaplex assay for detection of respiratory viruses in children. J Clin Microbiol. 2001;39:1696–701.PubMedCrossRef
7.
go back to reference Létant SE, Ortiz JI, Bentley Tammero LF, Birch JM, Derlet RW, Cohen S, Manning D, McBride MT. Multiplexed reverse transcriptase PCR assay for identification of viral respiratory pathogens at the point of care. J Clin Microbiol. 2007;45:3498–505.PubMedCrossRef Létant SE, Ortiz JI, Bentley Tammero LF, Birch JM, Derlet RW, Cohen S, Manning D, McBride MT. Multiplexed reverse transcriptase PCR assay for identification of viral respiratory pathogens at the point of care. J Clin Microbiol. 2007;45:3498–505.PubMedCrossRef
8.
go back to reference Nolte FS, Marshall DJ, Rasberry C, Schievelbein S, Banks GG, Storch GA, Arens MQ, Buller RS, Prudent JR. MultiCode-PLx system for multiplexed detection of seventeen respiratory viruses. J Clin Microbiol. 2007;45:2779–86.PubMedCrossRef Nolte FS, Marshall DJ, Rasberry C, Schievelbein S, Banks GG, Storch GA, Arens MQ, Buller RS, Prudent JR. MultiCode-PLx system for multiplexed detection of seventeen respiratory viruses. J Clin Microbiol. 2007;45:2779–86.PubMedCrossRef
9.
go back to reference Syrmis MW, Whiley DM, Thomas M, Mackay IM, Williamson J, Siebert DJ, Nissen MD, Sloots TP. A sensitive, specific, and cost-effective multiplex reverse transcriptase-PCR assay for the detection of seven common respiratory viruses in respiratory samples. J Mol Diagn. 2004;6:125–31.PubMedCrossRef Syrmis MW, Whiley DM, Thomas M, Mackay IM, Williamson J, Siebert DJ, Nissen MD, Sloots TP. A sensitive, specific, and cost-effective multiplex reverse transcriptase-PCR assay for the detection of seven common respiratory viruses in respiratory samples. J Mol Diagn. 2004;6:125–31.PubMedCrossRef
10.
go back to reference Templeton KE, Scheltinga SA, Beersma MF, Kroes AC, Claas EC. Rapid and sensitive method using multiplex real-time PCR for diagnosis of infections by influenza A and influenza B viruses, respiratory syncytial virus, and parainfluenza viruses 1, 2, 3, and 4. J Clin Microbiol. 2004;42:1564–9.PubMedCrossRef Templeton KE, Scheltinga SA, Beersma MF, Kroes AC, Claas EC. Rapid and sensitive method using multiplex real-time PCR for diagnosis of infections by influenza A and influenza B viruses, respiratory syncytial virus, and parainfluenza viruses 1, 2, 3, and 4. J Clin Microbiol. 2004;42:1564–9.PubMedCrossRef
11.
go back to reference Wasem S, Weichert S, Walther S, Weigl JA, Puppe W, Ihorst G, Schmitt HJ, Forster J. Lower respiratory tract disease in children: constant pathogens—constant management?! Klin Padiatr. 2008;220:291–5.PubMedCrossRef Wasem S, Weichert S, Walther S, Weigl JA, Puppe W, Ihorst G, Schmitt HJ, Forster J. Lower respiratory tract disease in children: constant pathogens—constant management?! Klin Padiatr. 2008;220:291–5.PubMedCrossRef
12.
go back to reference Weigl JA, Puppe W, Schmitt HJ. Incidence of respiratory syncytial virus-positive hospitalizations in Germany. Eur J Clin Microbiol Infect Dis. 2001;20:452–9.PubMed Weigl JA, Puppe W, Schmitt HJ. Incidence of respiratory syncytial virus-positive hospitalizations in Germany. Eur J Clin Microbiol Infect Dis. 2001;20:452–9.PubMed
13.
go back to reference Weigl JA, Puppe W, Schmitt HJ. The incidence of influenza-associated hospitalizations in children in Germany. Epidemiol Infect. 2002;129:525–33.PubMedCrossRef Weigl JA, Puppe W, Schmitt HJ. The incidence of influenza-associated hospitalizations in children in Germany. Epidemiol Infect. 2002;129:525–33.PubMedCrossRef
14.
go back to reference Weigl JA, Puppe W, Schmitt HJ. Can respiratory syncytial virus etiology be diagnosed clinically? A hospital-based case–control study in children under two years of age. Eur J Epidemiol. 2003;18:431–9.PubMedCrossRef Weigl JA, Puppe W, Schmitt HJ. Can respiratory syncytial virus etiology be diagnosed clinically? A hospital-based case–control study in children under two years of age. Eur J Epidemiol. 2003;18:431–9.PubMedCrossRef
15.
go back to reference Weigl JA, Puppe W, Schmitt HJ. Variables explaining the duration of hospitalization in children under two years of age admitted with acute airway infections: does respiratory syncytial virus have a direct impact? Klin Padiatr. 2004;216:7–15.PubMedCrossRef Weigl JA, Puppe W, Schmitt HJ. Variables explaining the duration of hospitalization in children under two years of age admitted with acute airway infections: does respiratory syncytial virus have a direct impact? Klin Padiatr. 2004;216:7–15.PubMedCrossRef
16.
go back to reference Weigl JA, Puppe W, Gröndahl B, Schmitt HJ. Epidemiological investigation of nine respiratory pathogens in hospitalized children in Germany using multiplex reverse-transcriptase polymerase chain reaction. Eur J Clin Microbiol Infect Dis. 2000;19:336–43.PubMedCrossRef Weigl JA, Puppe W, Gröndahl B, Schmitt HJ. Epidemiological investigation of nine respiratory pathogens in hospitalized children in Germany using multiplex reverse-transcriptase polymerase chain reaction. Eur J Clin Microbiol Infect Dis. 2000;19:336–43.PubMedCrossRef
17.
go back to reference Weigl JA, Puppe W, Rockahr S, Schmitt HJ. Burden of disease in hospitalized RSV-positive children in Germany. Klin Padiatr. 2002;214:334–42.PubMedCrossRef Weigl JA, Puppe W, Rockahr S, Schmitt HJ. Burden of disease in hospitalized RSV-positive children in Germany. Klin Padiatr. 2002;214:334–42.PubMedCrossRef
18.
go back to reference Weigl JA, Puppe W, Belke O, Neusüss J, Bagci F, Schmitt HJ. The descriptive epidemiology of severe lower respiratory tract infections in children in Kiel, Germany. Klin Padiatr. 2005;217:259–67.PubMedCrossRef Weigl JA, Puppe W, Belke O, Neusüss J, Bagci F, Schmitt HJ. The descriptive epidemiology of severe lower respiratory tract infections in children in Kiel, Germany. Klin Padiatr. 2005;217:259–67.PubMedCrossRef
19.
go back to reference Weigl JA, Puppe W, Belke O, Neusüss J, Bagci F, Schmitt HJ. Population-based incidence of severe pneumonia in children in Kiel, Germany. Klin Padiatr. 2005;217:211–9.PubMedCrossRef Weigl JA, Puppe W, Belke O, Neusüss J, Bagci F, Schmitt HJ. Population-based incidence of severe pneumonia in children in Kiel, Germany. Klin Padiatr. 2005;217:211–9.PubMedCrossRef
20.
go back to reference Weigl JA, Puppe W, Meyer CU, Berner R, Forster J, Schmitt HJ, Zepp F. Ten years’ experience with year-round active surveillance of up to 19 respiratory pathogens in children. Eur J Pediatr. 2007;166:957–66.PubMedCrossRef Weigl JA, Puppe W, Meyer CU, Berner R, Forster J, Schmitt HJ, Zepp F. Ten years’ experience with year-round active surveillance of up to 19 respiratory pathogens in children. Eur J Pediatr. 2007;166:957–66.PubMedCrossRef
21.
go back to reference Weigl JA, Puppe W, Meyer CU, Berner R, Forster J, Schmitt HJ, Zepp F. PID-ARI.net—a pediatric infectious diseases network on acute respiratory infections and the added value of a multilevel research network. Klin Padiatr. 2008;220:281–6.PubMedCrossRef Weigl JA, Puppe W, Meyer CU, Berner R, Forster J, Schmitt HJ, Zepp F. PID-ARI.net—a pediatric infectious diseases network on acute respiratory infections and the added value of a multilevel research network. Klin Padiatr. 2008;220:281–6.PubMedCrossRef
22.
go back to reference du Prel JB, Puppe W, Gröndahl B, Knuf M, Weigl JA, Schaaff F, Schmitt HJ. Are meteorological parameters associated with acute respiratory tract infections? Clin Infect Dis. 2009;49:861–8.PubMedCrossRef du Prel JB, Puppe W, Gröndahl B, Knuf M, Weigl JA, Schaaff F, Schmitt HJ. Are meteorological parameters associated with acute respiratory tract infections? Clin Infect Dis. 2009;49:861–8.PubMedCrossRef
23.
go back to reference Gröndahl B, Puppe W, Weigl J, Schmitt HJ. Comparison of the BD Directigen Flu A+B Kit and the Abbott TestPack RSV with a multiplex RT-PCR ELISA for rapid detection of influenza viruses and respiratory syncytial virus. Clin Microbiol Infect. 2005;11:848–50.PubMedCrossRef Gröndahl B, Puppe W, Weigl J, Schmitt HJ. Comparison of the BD Directigen Flu A+B Kit and the Abbott TestPack RSV with a multiplex RT-PCR ELISA for rapid detection of influenza viruses and respiratory syncytial virus. Clin Microbiol Infect. 2005;11:848–50.PubMedCrossRef
24.
go back to reference Huck B, Scharf G, Neumann-Häfelin D, Puppe W, Weigl J, Falcone V. Novel human metapneumovirus sublineage. Emerg Infect Dis. 2006;12:147–50.PubMedCrossRef Huck B, Scharf G, Neumann-Häfelin D, Puppe W, Weigl J, Falcone V. Novel human metapneumovirus sublineage. Emerg Infect Dis. 2006;12:147–50.PubMedCrossRef
25.
go back to reference Schützle H, Weigl J, Puppe W, Forster J, Berner R. Diagnostic performance of a rapid antigen test for RSV in comparison with a 19-valent multiplex RT-PCR ELISA in children with acute respiratory tract infections. Eur J Pediatr. 2008;167:745–9.PubMedCrossRef Schützle H, Weigl J, Puppe W, Forster J, Berner R. Diagnostic performance of a rapid antigen test for RSV in comparison with a 19-valent multiplex RT-PCR ELISA in children with acute respiratory tract infections. Eur J Pediatr. 2008;167:745–9.PubMedCrossRef
26.
go back to reference Toschke AM, Arenz S, von Kries R, Puppe W, Weigl JA, Höhle M, Heininger U. No temporal association between influenza outbreaks and invasive pneumococcal infections. Arch Dis Child. 2008;93:218–20.PubMedCrossRef Toschke AM, Arenz S, von Kries R, Puppe W, Weigl JA, Höhle M, Heininger U. No temporal association between influenza outbreaks and invasive pneumococcal infections. Arch Dis Child. 2008;93:218–20.PubMedCrossRef
27.
go back to reference Li Z, Jansen DL, Finn TM, Halperin SA, Kasina A, O’Connor SP, Aoyama T, Manclark CR, Brennan MJ. Identification of Bordetella pertussis infection by shared-primer PCR. J Clin Microbiol. 1994;32:783–9.PubMed Li Z, Jansen DL, Finn TM, Halperin SA, Kasina A, O’Connor SP, Aoyama T, Manclark CR, Brennan MJ. Identification of Bordetella pertussis infection by shared-primer PCR. J Clin Microbiol. 1994;32:783–9.PubMed
28.
go back to reference Pongthanapisith V, Sukasem C, Premchaiporn K, Srichantaratsamee C, Chantratita W. Clinical performance of three rapid diagnostic tests for influenza virus in nasopharyngeal specimens to detect novel swine-origin influenza viruses. Infection. 2011;39:105–11. Epub 2011 Mar 22. PubMed PMID: 21424855.PubMedCrossRef Pongthanapisith V, Sukasem C, Premchaiporn K, Srichantaratsamee C, Chantratita W. Clinical performance of three rapid diagnostic tests for influenza virus in nasopharyngeal specimens to detect novel swine-origin influenza viruses. Infection. 2011;39:105–11. Epub 2011 Mar 22. PubMed PMID: 21424855.PubMedCrossRef
29.
go back to reference Panning M, Eickmann M, Landt O, Monazahian M, Olschläger S, Baumgarte S, Reischl U, Wenzel JJ, Niller HH, Günther S, Hollmann B, Huzly D, Drexler JF, Helmer A, Becker S, Matz B, Eis-Hübinger A, Drosten C. Detection of influenza A(H1N1)v virus by real-time RT-PCR. Euro Surveill. 2009;14:pii=19329. Panning M, Eickmann M, Landt O, Monazahian M, Olschläger S, Baumgarte S, Reischl U, Wenzel JJ, Niller HH, Günther S, Hollmann B, Huzly D, Drexler JF, Helmer A, Becker S, Matz B, Eis-Hübinger A, Drosten C. Detection of influenza A(H1N1)v virus by real-time RT-PCR. Euro Surveill. 2009;14:pii=19329.
30.
go back to reference Engler HD, Selepak ST. Effect of centrifuging shell vials at 3,500 × g on detection of viruses in clinical specimens. J Clin Microbiol. 1994;32:1580–2.PubMed Engler HD, Selepak ST. Effect of centrifuging shell vials at 3,500 × g on detection of viruses in clinical specimens. J Clin Microbiol. 1994;32:1580–2.PubMed
31.
go back to reference Rothbarth PH, Groen J, Bohnen AM, de Groot R, Osterhaus AD. Influenza virus serology—a comparative study. J Virol Methods. 1999;78:163–9.PubMedCrossRef Rothbarth PH, Groen J, Bohnen AM, de Groot R, Osterhaus AD. Influenza virus serology—a comparative study. J Virol Methods. 1999;78:163–9.PubMedCrossRef
32.
go back to reference Dawson B, Trapp RG. Probability and related topics for making inferences about data. In: Dawson B, Trapp RG, editors. Basic and clinical biostatistics. 3rd ed. New York: Lange Medical Books/McGraw-Hill; 2001. p. 82–4. Dawson B, Trapp RG. Probability and related topics for making inferences about data. In: Dawson B, Trapp RG, editors. Basic and clinical biostatistics. 3rd ed. New York: Lange Medical Books/McGraw-Hill; 2001. p. 82–4.
33.
go back to reference Drosten C, Chiu LL, Panning M, Leong HN, Preiser W, Tam JS, Günther S, Kramme S, Emmerich P, Ng WL, Schmitz H, Koay ES. Evaluation of advanced reverse transcription-PCR assays and an alternative PCR target region for detection of severe acute respiratory syndrome-associated coronavirus. J Clin Microbiol. 2004;42:2043–7.PubMedCrossRef Drosten C, Chiu LL, Panning M, Leong HN, Preiser W, Tam JS, Günther S, Kramme S, Emmerich P, Ng WL, Schmitz H, Koay ES. Evaluation of advanced reverse transcription-PCR assays and an alternative PCR target region for detection of severe acute respiratory syndrome-associated coronavirus. J Clin Microbiol. 2004;42:2043–7.PubMedCrossRef
34.
go back to reference Drosten C, Günther S, Preiser W, van der Werf S, Brodt HR, Becker S, Rabenau H, Panning M, Kolesnikova L, Fouchier RA, Berger A, Burguière AM, Cinatl J, Eickmann M, Escriou N, Grywna K, Kramme S, Manuguerra JC, Müller S, Rickerts V, Stürmer M, Vieth S, Klenk HD, Osterhaus AD, Schmitz H, Doerr HW. Identification of a novel coronavirus in patients with severe acute respiratory syndrome. N Engl J Med. 2003;348:1967–76.PubMedCrossRef Drosten C, Günther S, Preiser W, van der Werf S, Brodt HR, Becker S, Rabenau H, Panning M, Kolesnikova L, Fouchier RA, Berger A, Burguière AM, Cinatl J, Eickmann M, Escriou N, Grywna K, Kramme S, Manuguerra JC, Müller S, Rickerts V, Stürmer M, Vieth S, Klenk HD, Osterhaus AD, Schmitz H, Doerr HW. Identification of a novel coronavirus in patients with severe acute respiratory syndrome. N Engl J Med. 2003;348:1967–76.PubMedCrossRef
35.
go back to reference Echevarría JE, Erdman DD, Swierkosz EM, Holloway BP, Anderson LJ. Simultaneous detection and identification of human parainfluenza viruses 1, 2, and 3 from clinical samples by multiplex PCR. J Clin Microbiol. 1998;36:1388–91.PubMed Echevarría JE, Erdman DD, Swierkosz EM, Holloway BP, Anderson LJ. Simultaneous detection and identification of human parainfluenza viruses 1, 2, and 3 from clinical samples by multiplex PCR. J Clin Microbiol. 1998;36:1388–91.PubMed
36.
go back to reference Arden KE, McErlean P, Nissen MD, Sloots TP, Mackay IM. Frequent detection of human rhinoviruses, paramyxoviruses, coronaviruses, and bocavirus during acute respiratory tract infections. J Med Virol. 2006;78:1232–40.PubMedCrossRef Arden KE, McErlean P, Nissen MD, Sloots TP, Mackay IM. Frequent detection of human rhinoviruses, paramyxoviruses, coronaviruses, and bocavirus during acute respiratory tract infections. J Med Virol. 2006;78:1232–40.PubMedCrossRef
37.
go back to reference Renwick N, Schweiger B, Kapoor V, Liu Z, Villari J, Bullmann R, Miething R, Briese T, Lipkin WI. A recently identified rhinovirus genotype is associated with severe respiratory-tract infection in children in Germany. J Infect Dis. 2007;196:1754–60.PubMedCrossRef Renwick N, Schweiger B, Kapoor V, Liu Z, Villari J, Bullmann R, Miething R, Briese T, Lipkin WI. A recently identified rhinovirus genotype is associated with severe respiratory-tract infection in children in Germany. J Infect Dis. 2007;196:1754–60.PubMedCrossRef
38.
go back to reference Benson R, Tondella ML, Bhatnagar J, Carvalho Mda G, Sampson JS, Talkington DF, Whitney AM, Mothershed E, McGee L, Carlone G, McClee V, Guarner J, Zaki S, Dejsiri S, Cronin K, Han J, Fields BS. Development and evaluation of a novel multiplex PCR technology for molecular differential detection of bacterial respiratory disease pathogens. J Clin Microbiol. 2008;46:2074–7.PubMedCrossRef Benson R, Tondella ML, Bhatnagar J, Carvalho Mda G, Sampson JS, Talkington DF, Whitney AM, Mothershed E, McGee L, Carlone G, McClee V, Guarner J, Zaki S, Dejsiri S, Cronin K, Han J, Fields BS. Development and evaluation of a novel multiplex PCR technology for molecular differential detection of bacterial respiratory disease pathogens. J Clin Microbiol. 2008;46:2074–7.PubMedCrossRef
39.
go back to reference Lam WY, Yeung AC, Tang JW, Ip M, Chan EW, Hui M, Chan PK. Rapid multiplex nested PCR for detection of respiratory viruses. J Clin Microbiol. 2007;45:3631–40.PubMedCrossRef Lam WY, Yeung AC, Tang JW, Ip M, Chan EW, Hui M, Chan PK. Rapid multiplex nested PCR for detection of respiratory viruses. J Clin Microbiol. 2007;45:3631–40.PubMedCrossRef
40.
go back to reference Marshall DJ, Reisdorf E, Harms G, Beaty E, Moser MJ, Lee WM, Gern JE, Nolte FS, Shult P, Prudent JR. Evaluation of a multiplexed PCR assay for detection of respiratory viral pathogens in a public health laboratory setting. J Clin Microbiol. 2007;45:3875–82.PubMedCrossRef Marshall DJ, Reisdorf E, Harms G, Beaty E, Moser MJ, Lee WM, Gern JE, Nolte FS, Shult P, Prudent JR. Evaluation of a multiplexed PCR assay for detection of respiratory viral pathogens in a public health laboratory setting. J Clin Microbiol. 2007;45:3875–82.PubMedCrossRef
41.
go back to reference Reijans M, Dingemans G, Klaassen CH, Meis JF, Keijdener J, Mulders B, Eadie K, van Leeuwen W, van Belkum A, Horrevorts AM, Simons G. RespiFinder: a new multiparameter test to differentially identify fifteen respiratory viruses. J Clin Microbiol. 2008;46:1232–40.PubMedCrossRef Reijans M, Dingemans G, Klaassen CH, Meis JF, Keijdener J, Mulders B, Eadie K, van Leeuwen W, van Belkum A, Horrevorts AM, Simons G. RespiFinder: a new multiparameter test to differentially identify fifteen respiratory viruses. J Clin Microbiol. 2008;46:1232–40.PubMedCrossRef
42.
go back to reference Hierholzer JC, Halonen PE, Dahlen PO, Bingham PG, McDonough MM. Detection of adenovirus in clinical specimens by polymerase chain reaction and liquid-phase hybridization quantitated by time-resolved fluorometry. J Clin Microbiol. 1993;31:1886–91.PubMed Hierholzer JC, Halonen PE, Dahlen PO, Bingham PG, McDonough MM. Detection of adenovirus in clinical specimens by polymerase chain reaction and liquid-phase hybridization quantitated by time-resolved fluorometry. J Clin Microbiol. 1993;31:1886–91.PubMed
43.
go back to reference Kösters K, Reischl U, Schmetz J, Riffelmann M, Wirsing von König CH. Real-time LightCycler PCR for detection and discrimination of Bordetella pertussis and Bordetella parapertussis. J Clin Microbiol. 2002;40:1719–22.PubMedCrossRef Kösters K, Reischl U, Schmetz J, Riffelmann M, Wirsing von König CH. Real-time LightCycler PCR for detection and discrimination of Bordetella pertussis and Bordetella parapertussis. J Clin Microbiol. 2002;40:1719–22.PubMedCrossRef
44.
go back to reference Reischl U, Lehn N, Sanden GN, Loeffelholz MJ. Real-time PCR assay targeting IS481 of Bordetella pertussis and molecular basis for detecting Bordetella holmesii. J Clin Microbiol. 2001;39:1963–6.PubMedCrossRef Reischl U, Lehn N, Sanden GN, Loeffelholz MJ. Real-time PCR assay targeting IS481 of Bordetella pertussis and molecular basis for detecting Bordetella holmesii. J Clin Microbiol. 2001;39:1963–6.PubMedCrossRef
45.
go back to reference Gaydos CA, Quinn TC, Eiden JJ. Identification of Chlamydia pneumoniae by DNA amplification of the 16S rRNA gene. J Clin Microbiol. 1992;30:796–800.PubMed Gaydos CA, Quinn TC, Eiden JJ. Identification of Chlamydia pneumoniae by DNA amplification of the 16S rRNA gene. J Clin Microbiol. 1992;30:796–800.PubMed
46.
go back to reference Stephensen CB, Casebolt DB, Gangopadhyay NN. Phylogenetic analysis of a highly conserved region of the polymerase gene from 11 coronaviruses and development of a consensus polymerase chain reaction assay. Virus Res. 1999;60:181–9.PubMedCrossRef Stephensen CB, Casebolt DB, Gangopadhyay NN. Phylogenetic analysis of a highly conserved region of the polymerase gene from 11 coronaviruses and development of a consensus polymerase chain reaction assay. Virus Res. 1999;60:181–9.PubMedCrossRef
47.
go back to reference Vabret A, Mouthon F, Mourez T, Gouarin S, Petitjean J, Freymuth F. Direct diagnosis of human respiratory coronaviruses 229E and OC43 by the polymerase chain reaction. J Virol Methods. 2001;97:59–66.PubMedCrossRef Vabret A, Mouthon F, Mourez T, Gouarin S, Petitjean J, Freymuth F. Direct diagnosis of human respiratory coronaviruses 229E and OC43 by the polymerase chain reaction. J Virol Methods. 2001;97:59–66.PubMedCrossRef
48.
go back to reference Rotbart HA, Sawyer MH, Fast S, Lewinski C, Murphy N, Keyser EF, Spadoro J, Kao SY, Loeffelholz M. Diagnosis of enteroviral meningitis by using PCR with a colorimetric microwell detection assay. J Clin Microbiol. 1994;32:2590–2.PubMed Rotbart HA, Sawyer MH, Fast S, Lewinski C, Murphy N, Keyser EF, Spadoro J, Kao SY, Loeffelholz M. Diagnosis of enteroviral meningitis by using PCR with a colorimetric microwell detection assay. J Clin Microbiol. 1994;32:2590–2.PubMed
49.
go back to reference Claas EC, Sprenger MJ, Kleter GE, van Beek R, Quint WG, Masurel N. Type-specific identification of influenza viruses A, B and C by the polymerase chain reaction. J Virol Methods. 1992;39:1–13.PubMedCrossRef Claas EC, Sprenger MJ, Kleter GE, van Beek R, Quint WG, Masurel N. Type-specific identification of influenza viruses A, B and C by the polymerase chain reaction. J Virol Methods. 1992;39:1–13.PubMedCrossRef
50.
go back to reference Alexiou-Daniel S, Stylianakis A, Papoutsi A, Zorbas I, Papa A, Lambropoulos AF, Antoniadis A. Application of polymerase chain reaction for detection of Legionella pneumophila in serum samples. Clin Microbiol Infect. 1998;4:144–8.PubMedCrossRef Alexiou-Daniel S, Stylianakis A, Papoutsi A, Zorbas I, Papa A, Lambropoulos AF, Antoniadis A. Application of polymerase chain reaction for detection of Legionella pneumophila in serum samples. Clin Microbiol Infect. 1998;4:144–8.PubMedCrossRef
51.
go back to reference van Kuppeveld FJ, van der Logt JT, Angulo AF, van Zoest MJ, Quint WG, Niesters HG, Galama JM, Melchers WJ. Genus- and species-specific identification of mycoplasmas by 16S rRNA amplification. Appl Environ Microbiol. 1992;58:2606–15.PubMed van Kuppeveld FJ, van der Logt JT, Angulo AF, van Zoest MJ, Quint WG, Niesters HG, Galama JM, Melchers WJ. Genus- and species-specific identification of mycoplasmas by 16S rRNA amplification. Appl Environ Microbiol. 1992;58:2606–15.PubMed
52.
go back to reference Maertzdorf J, Wang CK, Brown JB, Quinto JD, Chu M, de Graaf M, van den Hoogen BG, Spaete R, Osterhaus AD, Fouchier RA. Real-time reverse transcriptase PCR assay for detection of human metapneumoviruses from all known genetic lineages. J Clin Microbiol. 2004;42:981–6.PubMedCrossRef Maertzdorf J, Wang CK, Brown JB, Quinto JD, Chu M, de Graaf M, van den Hoogen BG, Spaete R, Osterhaus AD, Fouchier RA. Real-time reverse transcriptase PCR assay for detection of human metapneumoviruses from all known genetic lineages. J Clin Microbiol. 2004;42:981–6.PubMedCrossRef
53.
go back to reference Fan J, Henrickson KJ. Rapid diagnosis of human parainfluenza virus type 1 infection by quantitative reverse transcription-PCR-enzyme hybridization assay. J Clin Microbiol. 1996;34:1914–7.PubMed Fan J, Henrickson KJ. Rapid diagnosis of human parainfluenza virus type 1 infection by quantitative reverse transcription-PCR-enzyme hybridization assay. J Clin Microbiol. 1996;34:1914–7.PubMed
54.
go back to reference Yuasa T, Bando H, Kawano M, Tsurudome M, Nishio M, Kondo K, Komada H, Ito Y. Sequence analyses of the 3′ genome end and NP gene of human parainfluenza type 2 virus: sequence variation of the gene-starting signal and the conserved 3′ end. Virology. 1990;179:777–84.PubMedCrossRef Yuasa T, Bando H, Kawano M, Tsurudome M, Nishio M, Kondo K, Komada H, Ito Y. Sequence analyses of the 3′ genome end and NP gene of human parainfluenza type 2 virus: sequence variation of the gene-starting signal and the conserved 3′ end. Virology. 1990;179:777–84.PubMedCrossRef
55.
go back to reference Aguilar JC, Pérez-Breña MP, García ML, Cruz N, Erdman DD, Echevarría JE. Detection and identification of human parainfluenza viruses 1, 2, 3, and 4 in clinical samples of pediatric patients by multiplex reverse transcription-PCR. J Clin Microbiol. 2000;38:1191–5.PubMed Aguilar JC, Pérez-Breña MP, García ML, Cruz N, Erdman DD, Echevarría JE. Detection and identification of human parainfluenza viruses 1, 2, 3, and 4 in clinical samples of pediatric patients by multiplex reverse transcription-PCR. J Clin Microbiol. 2000;38:1191–5.PubMed
56.
go back to reference Andeweg AC, Bestebroer TM, Huybreghs M, Kimman TG, de Jong JC. Improved detection of rhinoviruses in clinical samples by using a newly developed nested reverse transcription-PCR assay. J Clin Microbiol. 1999;37:524–30.PubMed Andeweg AC, Bestebroer TM, Huybreghs M, Kimman TG, de Jong JC. Improved detection of rhinoviruses in clinical samples by using a newly developed nested reverse transcription-PCR assay. J Clin Microbiol. 1999;37:524–30.PubMed
57.
go back to reference Pitkäranta A, Arruda E, Malmberg H, Hayden FG. Detection of rhinovirus in sinus brushings of patients with acute community-acquired sinusitis by reverse transcription-PCR. J Clin Microbiol. 1997;35:1791–3.PubMed Pitkäranta A, Arruda E, Malmberg H, Hayden FG. Detection of rhinovirus in sinus brushings of patients with acute community-acquired sinusitis by reverse transcription-PCR. J Clin Microbiol. 1997;35:1791–3.PubMed
58.
go back to reference Leary TP, Erker JC, Chalmers ML, Wetzel JD, Desai SM, Mushahwar IK, Dermody TS. Detection of reovirus by reverse transcription-polymerase chain reaction using primers corresponding to conserved regions of the viral L1 genome segment. J Virol Methods. 2002;104:161–5.PubMedCrossRef Leary TP, Erker JC, Chalmers ML, Wetzel JD, Desai SM, Mushahwar IK, Dermody TS. Detection of reovirus by reverse transcription-polymerase chain reaction using primers corresponding to conserved regions of the viral L1 genome segment. J Virol Methods. 2002;104:161–5.PubMedCrossRef
59.
go back to reference Paton AW, Paton JC, Lawrence AJ, Goldwater PN, Harris RJ. Rapid detection of respiratory syncytial virus in nasopharyngeal aspirates by reverse transcription and polymerase chain reaction amplification. J Clin Microbiol. 1992;30:901–4.PubMed Paton AW, Paton JC, Lawrence AJ, Goldwater PN, Harris RJ. Rapid detection of respiratory syncytial virus in nasopharyngeal aspirates by reverse transcription and polymerase chain reaction amplification. J Clin Microbiol. 1992;30:901–4.PubMed
Metadata
Title
Validation of a multiplex reverse transcriptase PCR ELISA for the detection of 19 respiratory tract pathogens
Authors
W. Puppe
J. Weigl
B. Gröndahl
M. Knuf
S. Rockahr
P. von Bismarck
G. Aron
H. G. M. Niesters
A. D. M. E. Osterhaus
H.-J. Schmitt
Publication date
01-02-2013
Publisher
Springer-Verlag
Published in
Infection / Issue 1/2013
Print ISSN: 0300-8126
Electronic ISSN: 1439-0973
DOI
https://doi.org/10.1007/s15010-012-0298-6

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