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Clinical evaluation of multiplex RT-PCR assays for the detection of influenza A/B and respiratory syncytial virus using a high throughput system

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Clinical evaluation of multiplex RT-PCR assays for the detection of influenza A/B and respiratory syncytial virus using a high throughput system. / Eigner, Ulrich; Reucher, Svenja; Hefner, Nadine; Staffa-Peichl, Sandrine; Kolb, Melissa; Betz, Ulrike; Holfelder, Martin; Spier, Gene; Pfefferle, Susanne; Lütgehetmann, Marc.

In: J VIROL METHODS, Vol. 269, 01.04.2019, p. 49-54.

Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review

Harvard

Eigner, U, Reucher, S, Hefner, N, Staffa-Peichl, S, Kolb, M, Betz, U, Holfelder, M, Spier, G, Pfefferle, S & Lütgehetmann, M 2019, 'Clinical evaluation of multiplex RT-PCR assays for the detection of influenza A/B and respiratory syncytial virus using a high throughput system', J VIROL METHODS, vol. 269, pp. 49-54. https://doi.org/10.1016/j.jviromet.2019.03.015

APA

Eigner, U., Reucher, S., Hefner, N., Staffa-Peichl, S., Kolb, M., Betz, U., Holfelder, M., Spier, G., Pfefferle, S., & Lütgehetmann, M. (2019). Clinical evaluation of multiplex RT-PCR assays for the detection of influenza A/B and respiratory syncytial virus using a high throughput system. J VIROL METHODS, 269, 49-54. https://doi.org/10.1016/j.jviromet.2019.03.015

Vancouver

Eigner U, Reucher S, Hefner N, Staffa-Peichl S, Kolb M, Betz U et al. Clinical evaluation of multiplex RT-PCR assays for the detection of influenza A/B and respiratory syncytial virus using a high throughput system. J VIROL METHODS. 2019 Apr 1;269:49-54. https://doi.org/10.1016/j.jviromet.2019.03.015

Bibtex

@article{d7bd2cac42164638a17e4b491fefefa3,
title = "Clinical evaluation of multiplex RT-PCR assays for the detection of influenza A/B and respiratory syncytial virus using a high throughput system",
abstract = "BACKGROUND: Lower respiratory tract infections are a major threat to public health systems worldwide, with RSV and influenza being the main agents causing hospitalization. In outbreak situations, high-volume respiratory testing is needed. In this study, we evaluated the analytical and clinical performance of a pre-designed primer/probe set for the simultaneous multiplex detection of both viruses on a high-throughput platform, the cobas{\textregistered} 6800, using the {"}open channel{"} of the system for integration of lab-developed assays for the detection of influenza and RSV.RESULTS: Using the influenza/RSV qPCR Assay with swabs, LoD (95%) in TCID50/mL for influenza-A was 0.009, influenza-B 0.003, RSV-A 0.202, and RSV-B 0.009. Inter-run variability (3xLoD) was low (<1 Ct for all targets). Of 371 clinical respiratory specimens analyzed, results were concordant for 358 samples. The calculated sensitivity and specificity of the assay were 98.3% and 98.4% for Flu-A, 100% and 98.5% for Flu-B, and 98.6% and 99.7% for RSV. All quality assessment panel specimens (N = 63, including avian influenza strains) were correctly identified. None of the tested microorganisms showed cross-reactivity.CONCLUSION: Compared with CE-IVD assays, the assay evaluated here showed good analytical and clinical sensitivity and specificity with broad coverage of different virus strains. It offers high-throughput capacity with low hands-on time, facilitating the laboratory management of large respiratory outbreaks.",
author = "Ulrich Eigner and Svenja Reucher and Nadine Hefner and Sandrine Staffa-Peichl and Melissa Kolb and Ulrike Betz and Martin Holfelder and Gene Spier and Susanne Pfefferle and Marc L{\"u}tgehetmann",
note = "Copyright {\textcopyright} 2019. Published by Elsevier B.V.",
year = "2019",
month = apr,
day = "1",
doi = "10.1016/j.jviromet.2019.03.015",
language = "English",
volume = "269",
pages = "49--54",
journal = "J VIROL METHODS",
issn = "0166-0934",
publisher = "Elsevier",

}

RIS

TY - JOUR

T1 - Clinical evaluation of multiplex RT-PCR assays for the detection of influenza A/B and respiratory syncytial virus using a high throughput system

AU - Eigner, Ulrich

AU - Reucher, Svenja

AU - Hefner, Nadine

AU - Staffa-Peichl, Sandrine

AU - Kolb, Melissa

AU - Betz, Ulrike

AU - Holfelder, Martin

AU - Spier, Gene

AU - Pfefferle, Susanne

AU - Lütgehetmann, Marc

N1 - Copyright © 2019. Published by Elsevier B.V.

PY - 2019/4/1

Y1 - 2019/4/1

N2 - BACKGROUND: Lower respiratory tract infections are a major threat to public health systems worldwide, with RSV and influenza being the main agents causing hospitalization. In outbreak situations, high-volume respiratory testing is needed. In this study, we evaluated the analytical and clinical performance of a pre-designed primer/probe set for the simultaneous multiplex detection of both viruses on a high-throughput platform, the cobas® 6800, using the "open channel" of the system for integration of lab-developed assays for the detection of influenza and RSV.RESULTS: Using the influenza/RSV qPCR Assay with swabs, LoD (95%) in TCID50/mL for influenza-A was 0.009, influenza-B 0.003, RSV-A 0.202, and RSV-B 0.009. Inter-run variability (3xLoD) was low (<1 Ct for all targets). Of 371 clinical respiratory specimens analyzed, results were concordant for 358 samples. The calculated sensitivity and specificity of the assay were 98.3% and 98.4% for Flu-A, 100% and 98.5% for Flu-B, and 98.6% and 99.7% for RSV. All quality assessment panel specimens (N = 63, including avian influenza strains) were correctly identified. None of the tested microorganisms showed cross-reactivity.CONCLUSION: Compared with CE-IVD assays, the assay evaluated here showed good analytical and clinical sensitivity and specificity with broad coverage of different virus strains. It offers high-throughput capacity with low hands-on time, facilitating the laboratory management of large respiratory outbreaks.

AB - BACKGROUND: Lower respiratory tract infections are a major threat to public health systems worldwide, with RSV and influenza being the main agents causing hospitalization. In outbreak situations, high-volume respiratory testing is needed. In this study, we evaluated the analytical and clinical performance of a pre-designed primer/probe set for the simultaneous multiplex detection of both viruses on a high-throughput platform, the cobas® 6800, using the "open channel" of the system for integration of lab-developed assays for the detection of influenza and RSV.RESULTS: Using the influenza/RSV qPCR Assay with swabs, LoD (95%) in TCID50/mL for influenza-A was 0.009, influenza-B 0.003, RSV-A 0.202, and RSV-B 0.009. Inter-run variability (3xLoD) was low (<1 Ct for all targets). Of 371 clinical respiratory specimens analyzed, results were concordant for 358 samples. The calculated sensitivity and specificity of the assay were 98.3% and 98.4% for Flu-A, 100% and 98.5% for Flu-B, and 98.6% and 99.7% for RSV. All quality assessment panel specimens (N = 63, including avian influenza strains) were correctly identified. None of the tested microorganisms showed cross-reactivity.CONCLUSION: Compared with CE-IVD assays, the assay evaluated here showed good analytical and clinical sensitivity and specificity with broad coverage of different virus strains. It offers high-throughput capacity with low hands-on time, facilitating the laboratory management of large respiratory outbreaks.

U2 - 10.1016/j.jviromet.2019.03.015

DO - 10.1016/j.jviromet.2019.03.015

M3 - SCORING: Journal article

C2 - 30946852

VL - 269

SP - 49

EP - 54

JO - J VIROL METHODS

JF - J VIROL METHODS

SN - 0166-0934

ER -