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

Journal Article · Wed Apr 11 04:00:00 EDT 2007 · Journal of Clinical Microbiology

DOI:https://doi.org/10.1128/JCM.01712-07· OSTI ID:940886

Letant, S E; Ortiz, J I; Tammero, L; Birch, J M; Derlet, R W; Cohen, S; Manning, D; McBride, M T

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.

• Research Organization: Lawrence Livermore National Laboratory (LLNL), Livermore, CA
• Sponsoring Organization: USDOE
• DOE Contract Number: W-7405-ENG-48
• OSTI ID: 940886
• Report Number(s): UCRL-JRNL-230177
• Journal Information: Journal of Clinical Microbiology, Journal Name: Journal of Clinical Microbiology Vol. 45; ISSN JCMIDW; ISSN 0095-1137
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
ADENOVIRUS
DATA ANALYSIS
DETECTION
DIAGNOSIS
IMPLEMENTATION
INFLUENZA
PATHOGENS
PERFORMANCE
PROCESSING
SENSITIVITY
SPECIFICITY
TESTING