Development and Evaluation of an Enterovirus D68 Real-Time Reverse Transcriptase PCR Assay

We have developed and evaluated a real-time reverse transcriptase PCR (RT-PCR) assay for the detection of human enterovirus D68 (EV-D68) in clinical specimens. This assay was developed in response to the unprecedented 2014 nationwide EV-D68 outbreak in the United States associated with severe respir...

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Main Authors:	Wylie, Todd N., Wylie, Kristine M., Buller, Richard S., Cannella, Maria, Storch, Gregory A.
Format:	Online
Language:	English
Published:	American Society for Microbiology 2015
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4508392/

id	pubmed-4508392
recordtype	oai_dc
spelling	pubmed-45083922015-07-29 Development and Evaluation of an Enterovirus D68 Real-Time Reverse Transcriptase PCR Assay Wylie, Todd N. Wylie, Kristine M. Buller, Richard S. Cannella, Maria Storch, Gregory A. Virology We have developed and evaluated a real-time reverse transcriptase PCR (RT-PCR) assay for the detection of human enterovirus D68 (EV-D68) in clinical specimens. This assay was developed in response to the unprecedented 2014 nationwide EV-D68 outbreak in the United States associated with severe respiratory illness. As part of our evaluation of the outbreak, we sequenced and published the genome sequence of the EV-D68 virus circulating in St. Louis, MO. This sequence, along with other GenBank sequences from past EV-D68 occurrences, was used to computationally select a region of EV-D68 appropriate for targeting in a strain-specific RT-PCR assay. The RT-PCR assay amplifies a segment of the VP1 gene, with an analytic limit of detection of 4 copies per reaction, and it was more sensitive than commercially available assays that detect enteroviruses and rhinoviruses without distinguishing between the two, including three multiplex respiratory panels approved for clinical use by the FDA. The assay did not detect any other enteroviruses or rhinoviruses tested and did detect divergent strains of EV-D68, including the first EV-D68 strain (Fermon) identified in California in 1962. This assay should be useful for identifying and studying current and future outbreaks of EV-D68 viruses. American Society for Microbiology 2015-07-20 2015-08 /pmc/articles/PMC4508392/ /pubmed/26063859 http://dx.doi.org/10.1128/JCM.00923-15 Text en Copyright © 2015, Wylie et al. http://creativecommons.org/licenses/by-nc-sa/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-ShareAlike 3.0 Unported license (http://creativecommons.org/licenses/by-nc-sa/3.0/) , which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original author and source are credited.
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Wylie, Todd N. Wylie, Kristine M. Buller, Richard S. Cannella, Maria Storch, Gregory A.
spellingShingle	Wylie, Todd N. Wylie, Kristine M. Buller, Richard S. Cannella, Maria Storch, Gregory A. Development and Evaluation of an Enterovirus D68 Real-Time Reverse Transcriptase PCR Assay
author_facet	Wylie, Todd N. Wylie, Kristine M. Buller, Richard S. Cannella, Maria Storch, Gregory A.
author_sort	Wylie, Todd N.
title	Development and Evaluation of an Enterovirus D68 Real-Time Reverse Transcriptase PCR Assay
title_short	Development and Evaluation of an Enterovirus D68 Real-Time Reverse Transcriptase PCR Assay
title_full	Development and Evaluation of an Enterovirus D68 Real-Time Reverse Transcriptase PCR Assay
title_fullStr	Development and Evaluation of an Enterovirus D68 Real-Time Reverse Transcriptase PCR Assay
title_full_unstemmed	Development and Evaluation of an Enterovirus D68 Real-Time Reverse Transcriptase PCR Assay
title_sort	development and evaluation of an enterovirus d68 real-time reverse transcriptase pcr assay
description	We have developed and evaluated a real-time reverse transcriptase PCR (RT-PCR) assay for the detection of human enterovirus D68 (EV-D68) in clinical specimens. This assay was developed in response to the unprecedented 2014 nationwide EV-D68 outbreak in the United States associated with severe respiratory illness. As part of our evaluation of the outbreak, we sequenced and published the genome sequence of the EV-D68 virus circulating in St. Louis, MO. This sequence, along with other GenBank sequences from past EV-D68 occurrences, was used to computationally select a region of EV-D68 appropriate for targeting in a strain-specific RT-PCR assay. The RT-PCR assay amplifies a segment of the VP1 gene, with an analytic limit of detection of 4 copies per reaction, and it was more sensitive than commercially available assays that detect enteroviruses and rhinoviruses without distinguishing between the two, including three multiplex respiratory panels approved for clinical use by the FDA. The assay did not detect any other enteroviruses or rhinoviruses tested and did detect divergent strains of EV-D68, including the first EV-D68 strain (Fermon) identified in California in 1962. This assay should be useful for identifying and studying current and future outbreaks of EV-D68 viruses.
publisher	American Society for Microbiology
publishDate	2015
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4508392/
_version_	1613249799431127040

Development and Evaluation of an Enterovirus D68 Real-Time Reverse Transcriptase PCR Assay

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