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Title: A multiplexed reverse transcriptase PCR assay for identification of viral respiratory pathogens at point-of-care

Abstract

We have developed a nucleic acid-based assay that is rapid, sensitive, specific, and can be used for the simultaneous detection of 5 common human respiratory pathogens including influenza A, influenza B, parainfluenza type 1 and 3, respiratory syncytial virus, and adenovirus group B, C, and E. Typically, diagnosis on an un-extracted clinical sample can be provided in less than 3 hours, including sample collection, preparation, and processing, as well as data analysis. Such a multiplexed panel would enable rapid broad-spectrum pathogen testing on nasal swabs, and therefore allow implementation of infection control measures, and timely administration of antiviral therapies. This article presents a summary of the assay performance in terms of sensitivity and specificity. Limits of detection are provided for each targeted respiratory pathogen, and result comparisons are performed on clinical samples, our goal being to compare the sensitivity and specificity of the multiplexed assay to the combination of immunofluorescence and shell vial culture currently implemented at the UCDMC hospital. Overall, the use of the multiplexed RT-PCR assay reduced the rate of false negatives by 4% and reduced the rate of false positives by up to 10%. The assay correctly identified 99.3% of the clinical negatives, 97% of adenovirus, 95%more » of RSV, 92% of influenza B, and 77% of influenza A without any extraction performed on the clinical samples. The data also showed that extraction will be needed for parainfluenza virus, which was only identified correctly 24% of the time on un-extracted samples.« less

Authors:
; ; ; ; ; ; ;
Publication Date:
Research Org.:
Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
940886
Report Number(s):
UCRL-JRNL-230177
Journal ID: ISSN 0095-1137; JCMIDW; TRN: US200824%%382
DOE Contract Number:  
W-7405-ENG-48
Resource Type:
Journal Article
Resource Relation:
Journal Name: Journal of Clinical Microbiology, vol. 45, N/A, November 1, 2007, pp. 3498-3505; Journal Volume: 45
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; ADENOVIRUS; DATA ANALYSIS; DETECTION; DIAGNOSIS; IMPLEMENTATION; INFLUENZA; PATHOGENS; PERFORMANCE; PROCESSING; SENSITIVITY; SPECIFICITY; TESTING

Citation Formats

Letant, S E, .Ortiz, J I, Tammero, L, Birch, J M, Derlet, R W, Cohen, S, Manning, D, and McBride, M T. A multiplexed reverse transcriptase PCR assay for identification of viral respiratory pathogens at point-of-care. United States: N. p., 2007. Web. doi:10.1128/JCM.01712-07.
Letant, S E, .Ortiz, J I, Tammero, L, Birch, J M, Derlet, R W, Cohen, S, Manning, D, & McBride, M T. A multiplexed reverse transcriptase PCR assay for identification of viral respiratory pathogens at point-of-care. United States. doi:10.1128/JCM.01712-07.
Letant, S E, .Ortiz, J I, Tammero, L, Birch, J M, Derlet, R W, Cohen, S, Manning, D, and McBride, M T. Wed . "A multiplexed reverse transcriptase PCR assay for identification of viral respiratory pathogens at point-of-care". United States. doi:10.1128/JCM.01712-07. https://www.osti.gov/servlets/purl/940886.
@article{osti_940886,
title = {A multiplexed reverse transcriptase PCR assay for identification of viral respiratory pathogens at point-of-care},
author = {Letant, S E and .Ortiz, J I and Tammero, L and Birch, J M and Derlet, R W and Cohen, S and Manning, D and McBride, M T},
abstractNote = {We have developed a nucleic acid-based assay that is rapid, sensitive, specific, and can be used for the simultaneous detection of 5 common human respiratory pathogens including influenza A, influenza B, parainfluenza type 1 and 3, respiratory syncytial virus, and adenovirus group B, C, and E. Typically, diagnosis on an un-extracted clinical sample can be provided in less than 3 hours, including sample collection, preparation, and processing, as well as data analysis. Such a multiplexed panel would enable rapid broad-spectrum pathogen testing on nasal swabs, and therefore allow implementation of infection control measures, and timely administration of antiviral therapies. This article presents a summary of the assay performance in terms of sensitivity and specificity. Limits of detection are provided for each targeted respiratory pathogen, and result comparisons are performed on clinical samples, our goal being to compare the sensitivity and specificity of the multiplexed assay to the combination of immunofluorescence and shell vial culture currently implemented at the UCDMC hospital. Overall, the use of the multiplexed RT-PCR assay reduced the rate of false negatives by 4% and reduced the rate of false positives by up to 10%. The assay correctly identified 99.3% of the clinical negatives, 97% of adenovirus, 95% of RSV, 92% of influenza B, and 77% of influenza A without any extraction performed on the clinical samples. The data also showed that extraction will be needed for parainfluenza virus, which was only identified correctly 24% of the time on un-extracted samples.},
doi = {10.1128/JCM.01712-07},
journal = {Journal of Clinical Microbiology, vol. 45, N/A, November 1, 2007, pp. 3498-3505},
number = ,
volume = 45,
place = {United States},
year = {Wed Apr 11 00:00:00 EDT 2007},
month = {Wed Apr 11 00:00:00 EDT 2007}
}