Chemiluminescent assay for the detection of viral and plasmid DNA using digoxigenin-labeled probes.
A dot-blot hybridization immunoenzymatic assay with a chemiluminescent endpoint was developed for the rapid and sensitive detection of viral and plasmid DNAs. Digoxigenin-labeled probes were used to detect cytomegalovirus, parvovirus B19, and plasmid pBR328 DNAs. Hybridized probes were immunoenzymatically visualized by anti-digoxigenin Fab fragments labeled with alkaline phosphatase, and adamantyl 1,2-dioxetane phenyl phosphate was used as chemiluminescent substrate. Results were recorded by instant photographic films. The chemiluminescent hybridization assay was performed in about 8 hr and was able to detect as little as 50-10 fg of homologous target DNA.
Musiani
M
M
Istituto di Microbiologia, Università di Bologna, Italy.
Zerbini
M
M
Gibellini
D
D
Gentilomi
G
G
La Placa
M
M
Ferri
E
E
Girotti
S
S
D003587:Cytomegalovirus / Q000235:genetics; D015342:DNA Probes; D004279:DNA, Viral / Q000032:analysis*; D004076:Digoxigenin; D007124:Immunoenzyme Techniques; D008163:Luminescent Measurements; D009693:Nucleic Acid Hybridization; D010321:Parvoviridae / Q000235:genetics; D010957:Plasmids
D015342:DNA Probes; D004279:DNA, Viral; D004076:Digoxigenin