Evaluation and Verification of the Seeplex Diarrhea-V ACE Assay for Simultaneous Detection of Adenovirus, Rotavirus, and Norovirus Genogroups I and II in Clinical Stool Specimens

Evaluation and Verification of the Seeplex Diarrhea-V ACE Assay for Simultaneous Detection of Adenovirus, Rotavirus, and Norovirus Genogroups I and II in Clinical Stool Specimens

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Published in:Journal of Clinical Microbiology Vol. 49; no. 9; pp. 3154 - 3162
Main Authors: HIGGINS, Rachel R, BENIPRASHAD, Melissa, CARDONA, Mark, MASNEY, Steven, LOW, Donald E, GUBBAY, Jonathan B
Format: Journal Article
Language:English
Published: Washington, DC American Society for Microbiology 01-09-2011
Subjects:
Adenoviridae - classification
Adenoviridae - isolation & purification
Adolescent
Adult
Aged
Aged, 80 and over
Biological and medical sciences
Child
Child, Preschool
Feces - virology
Female
Fundamental and applied biological sciences. Psychology
Gastroenteritis - virology
Genotype
Humans
Infant
Male
Microbiology
Middle Aged
Miscellaneous
Multiplex Polymerase Chain Reaction - methods
Norovirus - classification
Norovirus - isolation & purification
Retrospective Studies
Reverse Transcriptase Polymerase Chain Reaction - methods
Rotavirus - classification
Rotavirus - isolation & purification
Sensitivity and Specificity
Virology
Virology - methods
Virus Diseases - diagnosis
Virus Diseases - virology
Young Adult
Virus
Adenoviridae
Norovirus
Rotavirus
Caliciviridae
Diarrhea
Digestive diseases
Intestinal disease
Feces
Detection
Clinical isolate
Reoviridae
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Abstract Article Usage Stats Services JCM Citing Articles Google Scholar PubMed Related Content Social Bookmarking CiteULike Delicious Digg Facebook Google+ Mendeley Reddit StumbleUpon Twitter current issue JCM About JCM Subscribers Authors Reviewers Advertisers Inquiries from the Press Permissions & Commercial Reprints ASM Journals Public Access Policy JCM RSS Feeds 1752 N Street N.W. • Washington DC 20036 202.737.3600 • 202.942.9355 fax • journals@asmusa.org Print ISSN: 0095-1137 Online ISSN: 1098-660X Copyright © 2014 by the American Society for Microbiology.   For an alternate route to JCM .asm.org, visit: JCM       
AbstractList ABSTRACT Acute viral gastroenteritis is an intestinal infection that can be caused by several different viruses. Here we describe the evaluation and verification of Seeplex Diarrhea-V ACE (Seeplex DV), a novel commercial multiplex reverse transcription-PCR (RT-PCR) assay that detects 5 diarrheal pathogens, including adenovirus, rotavirus, norovirus genogroup I (GI) and GII, and astrovirus. We describe a retrospective study of 200 clinical specimens of which 177 were stool specimens previously tested for the presence of gastrointestinal viruses by electron microscopy (EM) and/or real-time RT-PCR (rRT-PCR). The remaining 23 specimens comprised other human pathogens of viral or bacterial origin. Discordant norovirus GI and GII results were resolved using a commercial kit; discordant adenovirus and rotavirus results were resolved using a home brew multiplex rRT-PCR assay. Diagnostic sensitivities and specificities were calculated before and after discordant analysis. After discordant analysis, estimated diagnostic sensitivities were 100% for adenovirus, rotavirus, and norovirus GI and 97% for norovirus GII. Diagnostic specificities after discordant analysis were 100% for adenovirus, rotavirus, and norovirus GI and 99.4% for norovirus GII. The 95% limits of detection were 31, 10, 2, and 1 genome equivalent per reaction for adenovirus, rotavirus, and norovirus GI and GII, respectively. The results demonstrate that the Seeplex DV assay is sensitive, specific, convenient, and reliable for the simultaneous detection of several viral pathogens directly in specimens from patients with gastroenteritis. Importantly, this novel multiplex PCR assay enabled the identification of viral coinfections in 12 (6.8%) stool specimens.
Acute viral gastroenteritis is an intestinal infection that can be caused by several different viruses. Here we describe the evaluation and verification of Seeplex Diarrhea-V ACE (Seeplex DV), a novel commercial multiplex reverse transcription-PCR (RT-PCR) assay that detects 5 diarrheal pathogens, including adenovirus, rotavirus, norovirus genogroup I (GI) and GII, and astrovirus. We describe a retrospective study of 200 clinical specimens of which 177 were stool specimens previously tested for the presence of gastrointestinal viruses by electron microscopy (EM) and/or real-time RT-PCR (rRT-PCR). The remaining 23 specimens comprised other human pathogens of viral or bacterial origin. Discordant norovirus GI and GII results were resolved using a commercial kit; discordant adenovirus and rotavirus results were resolved using a home brew multiplex rRT-PCR assay. Diagnostic sensitivities and specificities were calculated before and after discordant analysis. After discordant analysis, estimated diagnostic sensitivities were 100% for adenovirus, rotavirus, and norovirus GI and 97% for norovirus GII. Diagnostic specificities after discordant analysis were 100% for adenovirus, rotavirus, and norovirus GI and 99.4% for norovirus GII. The 95% limits of detection were 31, 10, 2, and 1 genome equivalent per reaction for adenovirus, rotavirus, and norovirus GI and GII, respectively. The results demonstrate that the Seeplex DV assay is sensitive, specific, convenient, and reliable for the simultaneous detection of several viral pathogens directly in specimens from patients with gastroenteritis. Importantly, this novel multiplex PCR assay enabled the identification of viral coinfections in 12 (6.8%) stool specimens.
Article Usage Stats Services JCM Citing Articles Google Scholar PubMed Related Content Social Bookmarking CiteULike Delicious Digg Facebook Google+ Mendeley Reddit StumbleUpon Twitter current issue JCM About JCM Subscribers Authors Reviewers Advertisers Inquiries from the Press Permissions & Commercial Reprints ASM Journals Public Access Policy JCM RSS Feeds 1752 N Street N.W. • Washington DC 20036 202.737.3600 • 202.942.9355 fax • journals@asmusa.org Print ISSN: 0095-1137 Online ISSN: 1098-660X Copyright © 2014 by the American Society for Microbiology.   For an alternate route to JCM .asm.org, visit: JCM       
Author Donald E. Low
Steven Masney
Jonathan B. Gubbay
Rachel R. Higgins
Melissa Beniprashad
Mark Cardona
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  organization: Ontario Agency for Health Protection and Promotion, Toronto, Ontario, Canada
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  fullname: CARDONA, Mark
  organization: Ontario Agency for Health Protection and Promotion, Toronto, Ontario, Canada
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  surname: MASNEY
  fullname: MASNEY, Steven
  organization: Ontario Agency for Health Protection and Promotion, Toronto, Ontario, Canada
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  givenname: Donald E
  surname: LOW
  fullname: LOW, Donald E
  organization: Ontario Agency for Health Protection and Promotion, Toronto, Ontario, Canada
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  organization: Ontario Agency for Health Protection and Promotion, Toronto, Ontario, Canada
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Keywords Virus
Adenoviridae
Norovirus
Rotavirus
Caliciviridae
Diarrhea
Digestive diseases
Intestinal disease
Feces
Detection
Clinical isolate
Reoviridae
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Snippet Article Usage Stats Services JCM Citing Articles Google Scholar PubMed Related Content Social Bookmarking CiteULike Delicious Digg Facebook Google+ Mendeley...
Acute viral gastroenteritis is an intestinal infection that can be caused by several different viruses. Here we describe the evaluation and verification of...
ABSTRACT Acute viral gastroenteritis is an intestinal infection that can be caused by several different viruses. Here we describe the evaluation and...
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StartPage 3154
SubjectTerms Adenoviridae - classification
Adenoviridae - isolation & purification
Adolescent
Adult
Aged
Aged, 80 and over
Biological and medical sciences
Child
Child, Preschool
Feces - virology
Female
Fundamental and applied biological sciences. Psychology
Gastroenteritis - virology
Genotype
Humans
Infant
Male
Microbiology
Middle Aged
Miscellaneous
Multiplex Polymerase Chain Reaction - methods
Norovirus - classification
Norovirus - isolation & purification
Retrospective Studies
Reverse Transcriptase Polymerase Chain Reaction - methods
Rotavirus - classification
Rotavirus - isolation & purification
Sensitivity and Specificity
Virology
Virology - methods
Virus Diseases - diagnosis
Virus Diseases - virology
Young Adult
Title Evaluation and Verification of the Seeplex Diarrhea-V ACE Assay for Simultaneous Detection of Adenovirus, Rotavirus, and Norovirus Genogroups I and II in Clinical Stool Specimens
URI http://jcm.asm.org/content/49/9/3154.abstract
https://www.ncbi.nlm.nih.gov/pubmed/21775550
https://search.proquest.com/docview/886909092
https://pubmed.ncbi.nlm.nih.gov/PMC3165607
Volume 49
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