Evaluation of a Multiplexed PCR Assay for Detection of Respiratory Viral Pathogens in a Public Health Laboratory Setting

EraGen Biosciences, Inc., Madison, Wisconsin, Wisconsin State Laboratory of Hygiene, Department of Pediatrics and Medicine, University of Wisconsin - Madison, Madison, Wisconsin, Medical University of South Carolina, Charleston, South Carolina, Centrose, Madison Wisconsin

^* To whom correspondence should be addressed. Email: dmarshall{at}eragen.com.

	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 empower researchers and clinicians to perform multiplexed molecular diagnostics quickly and easily. To this end, a new multiplexed molecular test called 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 which included the use of a real-time influenza A/B RT-PCR test and routine viral culture identification. The standard testing algorithm identified 503 (73%) positive and 184 negative samples. Analyzing those same 687 samples, PLx-RVP detected 528 (77%) positives for one or more targets and 159 complete target negatives. There were 25 discordant results between the two systems; fourteen were positive for viruses not routinely tested for by the Wisconsin State Laboratory of Hygiene (WSLH) of which 13 were confirmed by real-time PCR. When results using the standard testing algorithm were considered "true positives", PLx-RVP showed an overall sensitivity of 99% and overall specificity of 87%. In total, PLx-RVP detected an additional 40 viral infections of which 11 were mixed infections.