Direct quantitative chemiluminescent assays for the detection of viral DNA

✍ Scribed by Stefano Girotti; Elida Ferri; Severino Ghini; Monica Musiani; Maria Luisa Zerbini; Davide Gibellini; Giovanna Gentilomi

• Publisher: Elsevier Science
• Year: 1991
• Tongue: English
• Weight: 681 KB
• Volume: 255
• Category: Article
• ISSN: 0003-2670
• DOI: 10.1016/0003-2670(91)80073-3

✦ Synopsis

A direct chenulummescent dot blot hybndlzatlon assay for the detectlon of B19 Parvovnus DNA IS described The hybndizatton test uses dlgomgenm-labelled probes which are immunoenzymatlcally revealed by anti-dlgomgenm Fab fragments conlugated with alkalme phosphatase CAP) or mth horaeradlsh peromdase WU') The chenulummescent sIgnal, obtamed from an enzyme-tnggerable dloxetane for AP or from the lummol-amplified reaction for HRP, IS directly measured by placmg the spot cut from the nylon solid support m a cuvette and msertmg It mto a lummometer Both the enzymatic systems usmg this direct chenulummescent detectton gave reproducible results for cahbratlon graphs and posltlve chmcal samples, wth higher reproduclbdlty and long hfetlme emlsslon (15 days) for the AP-dloxetane system Thus &ect method allowed up to 0 2 pg of homologous target DNA to be revealed The results obtamed wth the quantltatwe chemdummescent assay descrrbed were compared wth those obtamed m a hybndlzatlon assay wtth colonmetnc detectlon or photographlc chenulummescent detection and good agreement among the tests was found Keywords Chenulummescence, DNA, Dot blot hybndlzatton assays, Enzyme reations