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Evaluation of fast-track diagnostics and TaqMan array card real-time PCR assays for the detection of respiratory pathogens
Several commercial assays are now available to detect the nucleic acid of multiple respiratory pathogens from a single specimen. Head-to-head comparisons of such assays using a single set of standard specimens provide additional information about key assay parameters such as sensitivity, specificity...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Published by Elsevier B.V.
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7114243/ https://www.ncbi.nlm.nih.gov/pubmed/25448378 http://dx.doi.org/10.1016/j.mimet.2014.10.009 |
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author | Driscoll, Amanda J. Karron, Ruth A. Bhat, Niranjan Thumar, Bhagvanji Kodani, Maja Fields, Barry S. Whitney, Cynthia G. Levine, Orin S. O'Brien, Katherine L. Murdoch, David R. |
author_facet | Driscoll, Amanda J. Karron, Ruth A. Bhat, Niranjan Thumar, Bhagvanji Kodani, Maja Fields, Barry S. Whitney, Cynthia G. Levine, Orin S. O'Brien, Katherine L. Murdoch, David R. |
author_sort | Driscoll, Amanda J. |
collection | PubMed |
description | Several commercial assays are now available to detect the nucleic acid of multiple respiratory pathogens from a single specimen. Head-to-head comparisons of such assays using a single set of standard specimens provide additional information about key assay parameters such as sensitivity, specificity and lower limits of detection, and help to inform the decision regarding which method to use. We evaluated two real-time PCR platforms: the Fast-track Diagnostics® (FTD) multiplex respiratory panel and a TaqMan array card (TAC) for simultaneous uniplex detection of multiple respiratory pathogens. Two sets of samples were used to evaluate the assays. One set was created by spiking pooled nasal wash or phosphate buffered saline with specified volumes of known concentrations of virus and/or bacteria. Clinical nasal wash specimens from children with lower respiratory tract illness comprised the other set. Thirteen pathogen targets were compared between the two platforms. Testing with a validation panel of spiked samples revealed a sensitivity of 96.1% and 92.9% for the FTD and TAC assays, respectively. Specificity could not be reliably calculated due to a suspected contamination of the sample substrate. Inter-assay agreement was high (> 95%) for most targets. Previously untested clinical specimens tested by both assays revealed a high percent agreement (> 95%) for all except rhinovirus, enterovirus and Streptococcus pneumoniae. Limitations of this evaluation included extraction of the validation samples by two different methods and the evaluation of the assays in different laboratories. However, neither of these factors significantly impacted inter-assay agreement for these sets of samples, and it was demonstrated that both assays could reliably detect clinically relevant concentrations of bacterial and viral pathogens. |
format | Online Article Text |
id | pubmed-7114243 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Published by Elsevier B.V. |
record_format | MEDLINE/PubMed |
spelling | pubmed-71142432020-04-02 Evaluation of fast-track diagnostics and TaqMan array card real-time PCR assays for the detection of respiratory pathogens Driscoll, Amanda J. Karron, Ruth A. Bhat, Niranjan Thumar, Bhagvanji Kodani, Maja Fields, Barry S. Whitney, Cynthia G. Levine, Orin S. O'Brien, Katherine L. Murdoch, David R. J Microbiol Methods Article Several commercial assays are now available to detect the nucleic acid of multiple respiratory pathogens from a single specimen. Head-to-head comparisons of such assays using a single set of standard specimens provide additional information about key assay parameters such as sensitivity, specificity and lower limits of detection, and help to inform the decision regarding which method to use. We evaluated two real-time PCR platforms: the Fast-track Diagnostics® (FTD) multiplex respiratory panel and a TaqMan array card (TAC) for simultaneous uniplex detection of multiple respiratory pathogens. Two sets of samples were used to evaluate the assays. One set was created by spiking pooled nasal wash or phosphate buffered saline with specified volumes of known concentrations of virus and/or bacteria. Clinical nasal wash specimens from children with lower respiratory tract illness comprised the other set. Thirteen pathogen targets were compared between the two platforms. Testing with a validation panel of spiked samples revealed a sensitivity of 96.1% and 92.9% for the FTD and TAC assays, respectively. Specificity could not be reliably calculated due to a suspected contamination of the sample substrate. Inter-assay agreement was high (> 95%) for most targets. Previously untested clinical specimens tested by both assays revealed a high percent agreement (> 95%) for all except rhinovirus, enterovirus and Streptococcus pneumoniae. Limitations of this evaluation included extraction of the validation samples by two different methods and the evaluation of the assays in different laboratories. However, neither of these factors significantly impacted inter-assay agreement for these sets of samples, and it was demonstrated that both assays could reliably detect clinically relevant concentrations of bacterial and viral pathogens. Published by Elsevier B.V. 2014-12 2014-10-27 /pmc/articles/PMC7114243/ /pubmed/25448378 http://dx.doi.org/10.1016/j.mimet.2014.10.009 Text en Copyright © 2014 Published by Elsevier B.V. Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active. |
spellingShingle | Article Driscoll, Amanda J. Karron, Ruth A. Bhat, Niranjan Thumar, Bhagvanji Kodani, Maja Fields, Barry S. Whitney, Cynthia G. Levine, Orin S. O'Brien, Katherine L. Murdoch, David R. Evaluation of fast-track diagnostics and TaqMan array card real-time PCR assays for the detection of respiratory pathogens |
title | Evaluation of fast-track diagnostics and TaqMan array card real-time PCR assays for the detection of respiratory pathogens |
title_full | Evaluation of fast-track diagnostics and TaqMan array card real-time PCR assays for the detection of respiratory pathogens |
title_fullStr | Evaluation of fast-track diagnostics and TaqMan array card real-time PCR assays for the detection of respiratory pathogens |
title_full_unstemmed | Evaluation of fast-track diagnostics and TaqMan array card real-time PCR assays for the detection of respiratory pathogens |
title_short | Evaluation of fast-track diagnostics and TaqMan array card real-time PCR assays for the detection of respiratory pathogens |
title_sort | evaluation of fast-track diagnostics and taqman array card real-time pcr assays for the detection of respiratory pathogens |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7114243/ https://www.ncbi.nlm.nih.gov/pubmed/25448378 http://dx.doi.org/10.1016/j.mimet.2014.10.009 |
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