DETECTION AND QUANTITATION OF HUMAN RESPIRATORY SYNCYTIAL VIRUS (RSV)USING MINIGENOME CDNA AND A SINDBIS-VIRUS REPLICON - A PROTOTYPE ASSAY FOR NEGATIVE-STRAND RNA VIRUSES

We describe here a novel approach for detecting and quantitating human respiratory syncytial virus (RSV) based on expression of a reporter g ene from an RSV minigenome. BHK cells were cytoplasmically transformed with a noncytopathic Sindbis virus replicon expressing T7 RNA polymer ase. These cells were then cotransfected with T7 expression plasmids t hat contain the cDNA of an RSV minigenome and the genes for RSV nucleo capsid proteins N, Rand L The minigenome contains a reporter gene such as lacZ or CAT flanked by cia-acting RSV transcription signals. Subse quent infection of these cells with RSV resulted in a high level of re porter gene expression which could be inhibited by ribavirin. Mock-inf ected cells exhibited background levels of expression. This assay can be used to quantitate RSV and titer neutralizing antibody and may be a valuable tool for screening compounds for anti-RSV activity. it serve s as a prototype for other negative-strand RNA viruses. (C) 1998 Acade mic Press.