A colorimetric microtiter plate PCR system detects respiratory syncytial virus in nasal aspirates and discriminates subtypes A and B

We developed a colorimetric microtiter plate (MTP) PCR system for specific detection of the respiratory syncytial virus (RSV) nucleocapsid gene and di fferentiation of viral subtypes A and B. Of 47 pediatric nasal aspirate spe cimens, the sensitivity and specificity were 94.4% (17 of 18) and 100% (15 of 15), respectively, when compared with RSV cell culture isolation in HEp- 2 cells. An additional 14 specimens positive for adenoviruses, rhinoviruses , influenza, or parainfluenza viruses did not give positive reactions. PCR testing detected a mean of 0.15 (0.01 to 7.00) plaque-forming units of RSV virions. Inhibition of PCR amplification was detected in 33.3% (6/18) of un diluted specimens and could be avoided by a dilution (1:10) of extracted RN A without decreasing test sensitivity. RSV subtype, as determined by allele -specific probes, was identical to that determined by an immunofluorescence assay. These results indicate that the MTP PCR system provides a sensitive and specific test for clinical laboratory diagnosis and simultaneous subgr oup classification of RSV infection. (C) 1999 Elsevier Science Inc.