A simple vacuum dot-blot hybridisation assay for the detection of Drosophila A and C viruses in single Drosophila. 1992

P D Christian
Research School of Biological Sciences, Australian National University, Canberra.

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.

UI MeSH Term Description Entries
D007304 Insect Viruses Viruses infecting insects, the largest family being BACULOVIRIDAE. Insect Virus,Virus, Insect,Viruses, Insect
D009693 Nucleic Acid Hybridization Widely used technique which exploits the ability of complementary sequences in single-stranded DNAs or RNAs to pair with each other to form a double helix. Hybridization can take place between two complimentary DNA sequences, between a single-stranded DNA and a complementary RNA, or between two RNA sequences. The technique is used to detect and isolate specific sequences, measure homology, or define other characteristics of one or both strands. (Kendrew, Encyclopedia of Molecular Biology, 1994, p503) Genomic Hybridization,Acid Hybridization, Nucleic,Acid Hybridizations, Nucleic,Genomic Hybridizations,Hybridization, Genomic,Hybridization, Nucleic Acid,Hybridizations, Genomic,Hybridizations, Nucleic Acid,Nucleic Acid Hybridizations
D003001 Cloning, Molecular The insertion of recombinant DNA molecules from prokaryotic and/or eukaryotic sources into a replicating vehicle, such as a plasmid or virus vector, and the introduction of the resultant hybrid molecules into recipient cells without altering the viability of those cells. Molecular Cloning
D004279 DNA, Viral Deoxyribonucleic acid that makes up the genetic material of viruses. Viral DNA
D004331 Drosophila melanogaster A species of fruit fly frequently used in genetics because of the large size of its chromosomes. D. melanogaster,Drosophila melanogasters,melanogaster, Drosophila
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D012367 RNA, Viral Ribonucleic acid that makes up the genetic material of viruses. Viral RNA
D012680 Sensitivity and Specificity Binary classification measures to assess test results. Sensitivity or recall rate is the proportion of true positives. Specificity is the probability of correctly determining the absence of a condition. (From Last, Dictionary of Epidemiology, 2d ed) Specificity,Sensitivity,Specificity and Sensitivity

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