Comparison of electronic microarray to enzyme hybridization assay for multiplex reverse-transcriptase PCR detection of common respiratory viruses in children.

Abstract

A new assay, composed of the NGEN RVA (Nanogen, Inc., San Diego, CA; Prodesse, Inc., Waukesha, WI), which is a pair of analyte-specific reagents that allow the multiplex reverse transcriptase polymerase chain reaction (RT-PCR) and electronic microarray detection of influenza virus A and B, respiratory syncytial virus A and B, and human parainfluenza virus types 1, 2, and 3, was evaluated in comparison with the Hexaplex (Prodesse), a multiplex RT-PCR–enzyme hybridization assay. Comparisons included the detection of respiratory viruses from whole-virus stocks (ATCC) and from frozen pediatric respiratory specimens collected at Children's Hospital of Wisconsin between 1991 and October 1998. After the retesting of six indeterminants and 20 discrepants, overall agreement improved to 96% on the positives and 100% on negatives, with only eight specimens still discrepant. The RVA reagents allow a rapid, sensitive, and specific assay for detecting seven of the most common respiratory viruses in children.