title: 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.
authors.affiliation: EraGen Biosciences Inc, Madison, WI 53717, USA. dmarshall@eragen.com
authors.forename: David J
authors.initials: DJ
authors.lastname: Marshall
authors.forename: Erik
authors.initials: E
authors.lastname: Reisdorf
authors.forename: Gerda
authors.initials: G
authors.lastname: Harms
authors.forename: Edward
authors.initials: E
authors.lastname: Beaty
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Moser
authors.forename: Wai-Ming
authors.initials: WM
authors.lastname: Lee
authors.forename: James E
authors.initials: JE
authors.lastname: Gern
authors.forename: Frederick S
authors.initials: FS
authors.lastname: Nolte
authors.forename: Pete
authors.initials: P
authors.lastname: Shult
authors.forename: James R
authors.initials: JR
authors.lastname: Prudent
mesh_terms: D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012141:Respiratory Tract Infections / Q000821:virology*; D012680:Sensitivity and Specificity; D014776:Virus Cultivation; D014777:Virus Diseases / Q000175:diagnosis*; D014780:Viruses / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*