A sample-in-answer-out instrument for the detection of multiple respiratory pathogens in unprepared nasopharyngeal swab samples

John Regan, Sonia Létant, Kristl Adams, Nga Nguyen, Robert Derlet, Stuart H Cohen, Elizabeth Vitalis, Lance Tammero, Josue Ortiz, Mary McBride, James Birch

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Multiplex RT-PCR suspension array assays provide a powerful tool for identifying the causative agent(s) of respiratory infections. These assays are time consuming and laborious on a time-per-sample basis if only a few samples require processing. To address this shortcoming and provide an automated solution for fast detection and identification of viral pathogens, we developed the first automated multiplex RT-PCR suspension array instrument capable of handling unprepared clinical samples. The instrument requires less than 3 minutes of hands-on time for a result generated in ∼2.5 hours. In analytical studies, the instrument performed as well as manually performed assays. The performance of the instrument and loaded multiplex viral detection assay was then tested using unprepared nasopharyngeal samples. The instrument-performed assay detected 61 of 71 RSV positive samples, for a sensitivity of 85.9%. Adenovirus (n = 5) and influenza B (n = 3) were less prevalent in the sample set, but detected to similar levels, 80% and 75%, respectively. The same sample set was also tested using FDA approved immuno-assay rapid tests, and the instrument was found to be more sensitive than the rapid tests with the sole exception being influenza A (n = 16), which was poorly detected due to significant sequence mismatches between the influenza A primer/probe set included in the multiplex mixture and the circulating influenza A strains. Overall, these data demonstrate the developed prototype platform performs multiplex array assays as well as hand-performed assays, and that the instrument's sensitivity and specificity are dictated by the quality of the loaded multiplex assay.

Original languageEnglish (US)
Pages (from-to)2316-2322
Number of pages7
JournalAnalyst
Volume135
Issue number9
DOIs
StatePublished - Sep 2010

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Pathogens
Human Influenza
Assays
pathogen
assay
influenza
Multiplex Polymerase Chain Reaction
Suspensions
Adenoviridae
Respiratory Tract Infections
Hand
Sensitivity and Specificity
detection
immunoassay
probe
Processing

ASJC Scopus subject areas

  • Analytical Chemistry
  • Spectroscopy
  • Electrochemistry
  • Biochemistry
  • Environmental Chemistry

Cite this

A sample-in-answer-out instrument for the detection of multiple respiratory pathogens in unprepared nasopharyngeal swab samples. / Regan, John; Létant, Sonia; Adams, Kristl; Nguyen, Nga; Derlet, Robert; Cohen, Stuart H; Vitalis, Elizabeth; Tammero, Lance; Ortiz, Josue; McBride, Mary; Birch, James.

In: Analyst, Vol. 135, No. 9, 09.2010, p. 2316-2322.

Research output: Contribution to journalArticle

Regan, J, Létant, S, Adams, K, Nguyen, N, Derlet, R, Cohen, SH, Vitalis, E, Tammero, L, Ortiz, J, McBride, M & Birch, J 2010, 'A sample-in-answer-out instrument for the detection of multiple respiratory pathogens in unprepared nasopharyngeal swab samples', Analyst, vol. 135, no. 9, pp. 2316-2322. https://doi.org/10.1039/c0an00223b
Regan, John ; Létant, Sonia ; Adams, Kristl ; Nguyen, Nga ; Derlet, Robert ; Cohen, Stuart H ; Vitalis, Elizabeth ; Tammero, Lance ; Ortiz, Josue ; McBride, Mary ; Birch, James. / A sample-in-answer-out instrument for the detection of multiple respiratory pathogens in unprepared nasopharyngeal swab samples. In: Analyst. 2010 ; Vol. 135, No. 9. pp. 2316-2322.
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