Early detection of acute rhinovirus infections by a rapid reverse transcription-PCR assay

The development of a rhinovirus (RV)-RNA-specific reverse transcription (RT )-PCR assay is complicated by the close homology between the RV and enterov irus (EV) genomes in the highly conserved 5'-noncoding region, which is cho sen for primer design in most RT-PCR assays. We have developed a sensitive, rapid, and RV-specific nested RT-PCR assay and have used it to test nasoph aryngeal aspirates from 556 patients presenting with acute respiratory trac t infections. RV RNA was detected by nested RT-PCR not only in all of 52 sa mples that were RV positive by virus isolation methods but also in 124 of 3 67 samples that were negative by virus isolation methods and enzyme-linked immunosorbent assay (ELISA). In addition, in 23 of 137 samples that were po sitive for a different respiratory virus by virus isolation and/or ELISA, R V RNA was detected by RT-PCR. EVs, adenoviruses, respiratory syncytial viru ses, coronaviruses, and influenza and parainfluenza viruses, including clin ical isolates as well as stock viruses, were not amplified in our RV-specif ic RT-PCR assay, indicating that this assay was highly specific. The proces sing time was less than 2 days for the RT-PCR, as opposed to up to 2 weeks for virus isolation. These results indicate that nested RT-PCR is more sens itive than conventional methods for the detection of RV in patients experie ncing acute respiratory tract infections and represents the only reliable t ool for the early laboratory diagnosis of RV infections. This is especially important in light of new opportunities for therapy currently being develo ped.