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Human respiratory syncytial virus (RSV) continues to be the leading viral cause of severe acute lower respiratory tract disease in infants and children worldwide. A licensed vaccine or antiviral drug suitable for routine use remains unavailable. This invention relates to the use of murine pneumonia virus (MPV), a virus to which humans normally are not exposed to and that is not cross-protected with RSV, as a vector to express the RSV fusion (F) glycoprotein as an RSV vaccine candidate. The RSV F ORF was codon optimized.

Human Enterovirus D68 (EV-D68) is a non-polio enterovirus that can cause mild to severe respiratory illness, especially in infants and children with asthma. Since its identification, every year EV-D68 has been detected sporadically throughout the world. The US experienced a nationwide outbreak of EV-D68 associated with a particularly severe respiratory illness from mid-August to early November 2014, with 1,153 confirmed cases in 49 states and the District of Columbia. Although various established detection methods are available for EV-D68, enteroviruses evolve rapidly.

Ebola virus surveillance in wildlife vectors of infection transmission to humans.

Pneumonia virus of mice expression vector PSynK-PVMJ3666.

Streptococcus pneumoniae, or pneumococcus, is a type of bacteria that causes pneumococcal disease. Pneumococcal infections can range from ear and sinus infections to pneumonia and bloodstream infections. Children younger than 2 years old and adults 65 years or older are among those most at risk for disease.

Mycoplasma pneumoniae (M. pneumonia) can cause several different types of infection including chest colds and pneumonia. M. pneumoniae is a leading cause of community-acquired pneumonia. People of all ages are at risk for getting M. pneumonia infection, but it is most common among young adults and school-aged children. Current methods of detecting this agent are laborious and time consuming, so testing is not usually performed. However, knowing whether someone has M. pneumoniae infection is important for choosing the right antibiotic for treatment.

Ebola virus (EBOV) hemorrhagic fever is one of the most lethal viral infections and lacks a licensed vaccine. EBOV is transmitted by contact with body fluids from infected individuals including droplets or aerosols. Aerosolized EBOV could also be exploited for intentional virus spread. Therefore, vaccines that protect against mucosal and systemic exposure are needed.

The inventors have created a reverse genetics system for RSV strain B1 of antigenic subgroup B encoding a replication-competent recombinant RSV that contains a codon-optimized G ORF and expresses enhanced green fluorescence protein (GFP). There are two antigenic subgroups of RSV, subgroups A and B, and most of the available information and reagents are for subgroup A. Immunity against either subgroup has reduced effectiveness in restricting the heterologous subgroup, suggesting that an effective RSV vaccine might need to contain both subgroups.

This technology provides a new set of hybridoma cell lines each expressing a single monoclonal antibody against human respiratory syncytial virus (RSV) nonstructural protein 1 (NS1). These antibodies have variously been shown to detect NS1 protein in an enzyme-linked immunosorbent assay (ELISA), Western blot assay, immunofluorescence microscopy of paraformaldehyde-fixed cells, and flow cytometry. The various antibodies can vary in their efficiency in each of these assays.