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

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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 ... continued below

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Letant, S. E.; Ortiz, J. I.; Tammero, L.; Birch, J. M.; Derlet, R. W.; Cohen, S. et al. April 11, 2007.

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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.

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PDF-file: 34 pages; size: 0.4 Mbytes

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  • Journal Name: Journal of Clinical Microbiology, vol. 45, N/A, November 1, 2007, pp. 3498-3505; Journal Volume: 45

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  • Report No.: UCRL-JRNL-230177
  • Grant Number: W-7405-ENG-48
  • Office of Scientific & Technical Information Report Number: 940886
  • Archival Resource Key: ark:/67531/metadc893363

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Office of Scientific & Technical Information Technical Reports

Reports, articles and other documents harvested from the Office of Scientific and Technical Information.

Office of Scientific and Technical Information (OSTI) is the Department of Energy (DOE) office that collects, preserves, and disseminates DOE-sponsored research and development (R&D) results that are the outcomes of R&D projects or other funded activities at DOE labs and facilities nationwide and grantees at universities and other institutions.

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  • April 11, 2007

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  • Sept. 27, 2016, 1:39 a.m.

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  • July 13, 2017, 3:32 p.m.

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Letant, S. E.; Ortiz, J. I.; Tammero, L.; Birch, J. M.; Derlet, R. W.; Cohen, S. et al. A multiplexed reverse transcriptase PCR assay for identification of viral respiratory pathogens at point-of-care, article, April 11, 2007; Livermore, California. (digital.library.unt.edu/ark:/67531/metadc893363/: accessed November 24, 2017), University of North Texas Libraries, Digital Library, digital.library.unt.edu; crediting UNT Libraries Government Documents Department.