Comparison of a multiplex reverse transcription-PCR-enzyme hybridization assay with conventional viral culture and immunofluorescence techniques for the detection of seven viral respiratory pathogens

A multiplex reverse transcription-PCR-enzyme hybridization assay (RT-PCR-EH A; Hexaplex; Prodesse Inc., Waukesha, Wis.) was used for the simultaneous d etection of human parainfluenza virus types 1, 2, and 3, influenza virus ty pes A and B, and respiratory syncytial virus types A and B. One hundred for ty-three respiratory specimens from 126 patients were analyzed by RT-PCR-ER A, and the results were compared to those obtained by conventional viral cu lture and immunofluorescence (IF) methods. RT-PCR-EHA proved to be positive for 17 of 143 (11.9%) specimens, whereas 8 of 143 (5.6%) samples were posi tive by viral culture and/or IF. Eight samples were positive by both RT-PCR -ERA and conventional methods, while nine samples were RT-PCR-ERA positive and viral culture and IF negative. Eight of the nine samples with discordan t results were then independently tested by a different multiplex RT-PCR as say for influenza virus types A and B, and all eight proved to be positive. In comparison to viral culture and IF methods, RT-PCR-EHA gave a sensitivi ty and a specificity of 100 and 93%, respectively. Since RT-PCR-EHA was abl e to detect more positive samples, which would otherwise have been missed b y routine methods, we suggest that this multiplex RT-PCR-ERA provides a hig hly sensitive and specific means of diagnostic detection of major respirato ry viruses.