Evaluation of three commercial multiplex assays for the detection of respiratory viral infections

- Comparison of Dual Priming Oligonucleotide vs. Liquid Bead Suspension Array vs. Tagged Oligo Cleavage Extension chemistry for the detection of respiratory viruses from adult and pediatric populations.
- Co-infections are detected frequently with these multiplex assays, especially in children, that it may impact current policies to cohort patients based on the result of a single virus.
- All 3 commercial assays are based on different chemistries and are statistically non-inferior to each other in terms of sensitivity and specificity, both on prospective and retrospective samples.
- Choice of assay by a clinical lab depends on volumes, cost, and work-flow considerations.

BackgroundTimely identification of respiratory virus infection is essential to mitigate inappropriate antibiotic use and to implement appropriate treatment and/or infection control procedures. As such, multiplexed PCR assays have become standard in many virology laboratories.ObjectivesTo compare the Seeplex RV15 (test of record) with two newer generation multiplex assays, the Anyplex II RV16 and the xTAG respiratory virus panels.Study designTwo hundred and three retrospective and 36 prospective respiratory samples were tested by all three assays. Samples were deemed to be positive if they tested positive for a virus by at least two of the three respective assays. Negative samples also had to test negative by at least two of the three assays. Inconclusive samples were those that showed band signal intensity between 0 and 100 on the RV15, but had not been previously tested on the RV16 or xTAG.Results and conclusionsOverall sensitivity and specificity of all three assays were similar (∼85% and 100%, respectively). Given each assay can identify multiple different viruses, the targets reported by one assay did not always agree with each target from another assay. Partial discordant rates were 47% and 21% for positive and negative samples, respectively. These higher than expected partial discordant rates may be due to primer or chemistry differences amongst the three multiplex assays.