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Evaluation of a multiplexed PCR assay for detection of respiratory viral pathogens in a public health laboratory setting.


ABSTRACT: There are numerous viral and bacterial causes of respiratory disease. To enable rapid and sensitive detection of even the most prevalent causes, there is a need for more-simplified testing systems that enable researchers and clinicians to perform multiplexed molecular diagnostics quickly and easily. To this end, a new multiplexed molecular test called the MultiCode-PLx respiratory virus panel (PLx-RVP) was developed and then implemented in a public health laboratory setting. A total of 687 respiratory samples were analyzed for the presence of 17 viruses that commonly cause respiratory disease. As a comparator, the samples were also tested using a standard testing algorithm that included the use of a real-time influenza virus A and B reverse transcription-PCR test and routine viral culture identification. The standard testing algorithm identified 503 (73%) samples as positive and 184 as negative. Analyzing the same 687 samples, the PLx-RVP assay detected one or more targets in 528 (77%) samples and found 159 samples negative for all targets. There were 25 discordant results between the two systems; 14 samples were positive for viruses not routinely tested for by the Wisconsin State Laboratory of Hygiene, and 13 of these were confirmed by real-time PCR. When the results of the standard testing algorithm were considered "true positives," the PLx-RVP assay showed an overall sensitivity of 99% and an overall specificity of 87%. In total, the PLx-RVP assay detected an additional 40 viral infections, of which 11 were mixed infections.

SUBMITTER: Marshall DJ 

PROVIDER: S-EPMC2168541 | biostudies-literature | 2007 Dec

REPOSITORIES: biostudies-literature

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Evaluation of a multiplexed PCR assay for detection of respiratory viral pathogens in a public health laboratory setting.

Marshall David J DJ   Reisdorf Erik E   Harms Gerda G   Beaty Edward E   Moser Michael J MJ   Lee Wai-Ming WM   Gern James E JE   Nolte Frederick S FS   Shult Pete P   Prudent James R JR  

Journal of clinical microbiology 20071010 12


There are numerous viral and bacterial causes of respiratory disease. To enable rapid and sensitive detection of even the most prevalent causes, there is a need for more-simplified testing systems that enable researchers and clinicians to perform multiplexed molecular diagnostics quickly and easily. To this end, a new multiplexed molecular test called the MultiCode-PLx respiratory virus panel (PLx-RVP) was developed and then implemented in a public health laboratory setting. A total of 687 respi  ...[more]

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