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Journal of Clinical Microbiology, December 2007, p. 3875-3882, Vol. 45, No. 12
0095-1137/07/$08.00+0     doi:10.1128/JCM.00838-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Evaluation of a Multiplexed PCR Assay for Detection of Respiratory Viral Pathogens in a Public Health Laboratory Setting{triangledown} ,{dagger}

David J. Marshall,1* Erik Reisdorf,2 Gerda Harms,1 Edward Beaty,1 Michael J. Moser,1 Wai-Ming Lee,3 James E. Gern,3 Frederick S. Nolte,4 Pete Shult,2 and James R. Prudent5

EraGen Biosciences, Inc,1 Wisconsin State Laboratory of Hygiene,2 Department of Pediatrics and Medicine, University of Wisconsin—Madison,3 Madison, Wisconsin,5 Medical University of South Carolina, Charleston, South Carolina; and Centrose, Madison, Wisconsin4

Received 19 April 2007/ Returned for modification 24 May 2007/ Accepted 1 October 2007

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.

* Corresponding author. Mailing address: EraGen Biosciences, Inc., 918 Deming Way, Madison, WI 53717. Phone: (608) 662-9000. Fax: (608) 662-9003. E-mail: dmarshall{at}eragen.com

{triangledown} Published ahead of print on 10 October 2007.

{dagger} Supplemental material for this article may be found at http://jsm.asm.org/.

Journal of Clinical Microbiology, December 2007, p. 3875-3882, Vol. 45, No. 12
0095-1137/07/$08.00+0     doi:10.1128/JCM.00838-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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