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Performance characteristics of a quantitative PCR assay on repository stool specimens and smeared filter-paper cards

Authors
  • Tisdale, Michele D.1, 2, 3, 4
  • Mitra, Indrani1, 3
  • McCoy, Andrea J.5
  • Simons, Mark P.5
  • Reynolds, Nathanael D.5
  • Swierczewski, Brett E.6
  • Liu, Jie7
  • Houpt, Eric R.7
  • Fraser, Jamie A.1, 3
  • Riddle, Mark S.1
  • Tribble, David R.1
  • Lalani, Tahaniyat1, 2, 3
  • 1 Uniformed Services University of the Health Sciences, Bethesda, MD, USA , Bethesda (United States)
  • 2 Naval Medical Center Portsmouth, Portsmouth, VA, USA , Portsmouth (United States)
  • 3 The Henry M Jackson Foundation for the Advancement of Military Medicine, Inc, Bethesda, MD, USA , Bethesda (United States)
  • 4 Infectious Disease & Travel Clinic, Building 3, 1st Floor, Naval Medical Center Portsmouth, Portsmouth, VA, 23708, USA , Portsmouth (United States)
  • 5 Naval Medical Research Unit-6, Lima, Peru , Lima (Peru)
  • 6 Armed Forces Research Institute of Medical Sciences, Bangkok, Thailand , Bangkok (Thailand)
  • 7 University of Virginia, Charlottesville, VA, USA , Charlottesville (United States)
Type
Published Article
Journal
BMC Research Notes
Publisher
Springer (Biomed Central Ltd.)
Publication Date
Oct 30, 2020
Volume
13
Issue
1
Identifiers
DOI: 10.1186/s13104-020-05340-7
Source
Springer Nature
Keywords
License
Green

Abstract

ObjectiveStool repositories are a valuable resource for retrospective analyses including quantitative PCR assays to distinguish between asymptomatic shedding and clinical disease. The suitability of archival specimens for this purpose is unclear and requires assessment. We conducted a pilot study to evaluate pathogen detection by TaqMan Array Card (TAC) in travelers’ diarrhea (TD) stool specimens stored for 1–13 years, as well as the impact of transporting specimens on Whatman FTA Elute cards (FTA Cards) on detection.ResultsThe positive percent agreement (PPA) for TAC on stool vs. microbiologic testing was lower than our a priori PPA estimate of 80% for most pathogens: Shigella spp. (100% [95%CI 69–100%]), enterotoxigenic E coli (ETEC) (63% [95%CI 49–75%]), Campylobacter spp. (66% [95%CI 43–85%]) and Norovirus (37% [95%CI 16–61%]). Use of the FTA card resulted in a further reduction of PPA. Our findings suggest that archival specimens may lead to insensitive detection on quantitative PCR assays due to degradation of nucleic acid with prolonged storage, although our limited sample size precluded us from evaluating the impact of storage duration on nucleic acid yield. Additional studies are needed to understand the impact of storage duration on quantitative PCR data.

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