A simple vacuum dot-blot hybridisation assay for the detection of Drosophila A and C viruses in single Drosophila.
Specific cDNA clones were constructed from the single stranded RNA genome of Australian isolates of both Drosophila A and C viruses. These clones were used to develop a nucleic acid hybridisation assay capable of detecting reliably 3.9 ng of DAV and 19.3 ng of DCV virus particles, respectively. The sensitivity of the assays were largely unaffected by soluble host material. Single Drosophila naturally infected or artificially inoculated with DAV or DCV were found to contain in excess of 130 ng of virus. The results presented here demonstrate that the vacuum dot-blotting protocol and the hybridisation assays developed are capable of detecting DAV and DCV in single Drosophila and may therefore be applied to the study of DAV and DCV in natural Drosophila communities.
Christian
P D
PD
Research School of Biological Sciences, Australian National University, Canberra.
D000818:Animals; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000235:genetics; D004331:Drosophila melanogaster / Q000382:microbiology*; D007304:Insect Viruses / Q000235:genetics / Q000302:isolation & purification*; D009693:Nucleic Acid Hybridization; D012367:RNA, Viral / Q000235:genetics*; D012680:Sensitivity and Specificity
D004279:DNA, Viral; D012367:RNA, Viral