Chemiluminescent assay for the detection of viral and plasmid DNA using digoxigenin-labeled probes

• Published in: Analytical biochemistry Vol. 194; no. 2; pp. 394 - 398
• Main Authors: Musiani, M., Zerbini, M., Gibellini, D., Gentilomi, G., La Placa, M., Ferri, E., Girotti, S.
• Format: Journal Article
• Language: English
• Published: San Diego, CA Elsevier Inc 01.05.1991; Elsevier
• Subjects: Analytical, structural and metabolic biochemistry; Biological and medical sciences; Cytomegalovirus - genetics; Digoxigenin; DNA Probes; Dna, deoxyribonucleoproteins; DNA, Viral - analysis; Fundamental and applied biological sciences. Psychology; Immunoenzyme Techniques; Luminescent Measurements; Nucleic Acid Hybridization; Nucleic acids; Parvoviridae - genetics; parvovirus B19; Plasmids; Plasmid; Virus; DNA; Blotting assay; Chemiluminescence; Molecular probe; Chemical labelling; Detection; Enzyme immunoassay; Nucleic acid
• ISSN: 0003-2697; 1096-0309
• DOI: 10.1016/0003-2697(91)90247-Q

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 immunoenzy-matically 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.