Development and Application of Nucleic Acid Hybridization Techniques to Arbovirus Surveillance and Diagnosis

摘要

Sandwich hybridization techniques have been developed for detection of dengue virus RNA. Capture and detector probe constructs have been prepared in M13 and in vitro transcription plasmids, respectively. In preliminary trials to determine optimal concentrations of catcher and detector probe, nanogram levels of analyte RNA were readily detected. Type and group specific hybridization probes for detection of California group virus RNA have been developed. The type specific probe hybridizes to snowshoe hare virus infected cells but not to La Crosse infected cell. The probe is a cDNA of 2161 bp portion of the middle RNA segment. The group specific probe hybridizes to cells infected with any of the California group viruses. The probe is a cDNA of the small RNA segment of La Crosse virus.