title: Digoxygenin-labelled DNA-probe: a rapid non-radioactive method for hepatitis B virus DNA detection in serum.
abstract: The sensitivity and specificity of two non-radioactive spot hybridization assays for hepatitis B virus DNA (HBV-DNA) using biotin and digoxygenin-labelled DNA probes were investigated in parallel in 122 serum samples from patients with chronic hepatitis B and 50 controls. The results were compared with an isotopic technique using a 32P-labelled probe. HBV-DNA was detected in 56 (80%) out of 70 hepatitis B "e" antigen (HBeAg)-positive cases and in 4 (8%) out of 52 antibody to hepatitis B "e" antigen (anti-HBe)-positive cases using the digoxygenin or 32P-labelled probes. No false positives were found with either method. Using the biotin-labelled probe, 16% of sera gave discordant results, which were considered to be false positive. The time required for detection of serum HBV-DNA was 2 hours for the non-radioactive probes and 16 hours for the isotopic probes. This study suggests that the digoxygenin-labelled probe for detection of HBV-DNA is the most rapid and sensitive method for routine diagnosis of viral replication in clinical laboratories.
authors.affiliation: Department of Internal Medicine, Hospital Valle Hebrón, Universidad Autónoma, Barcelona, Spain.
authors.forename: M
authors.initials: M
authors.lastname: Buti
authors.forename: R
authors.initials: R
authors.lastname: Jardi
authors.forename: F
authors.initials: F
authors.lastname: Rodriguez-Frias
authors.forename: J A
authors.initials: JA
authors.lastname: Arranz
authors.forename: J M
authors.initials: JM
authors.lastname: Casacuberta
authors.forename: B
authors.initials: B
authors.lastname: Sansegundo
authors.forename: P
authors.initials: P
authors.lastname: Puigdomenech
authors.forename: R
authors.initials: R
authors.lastname: Esteban
authors.forename: J
authors.initials: J
authors.lastname: Guardia
mesh_terms: D000410:Alanine Transaminase / Q000097:blood; D002908:Chronic Disease; D015342:DNA Probes; D004279:DNA, Viral / Q000097:blood*; D004076:Digoxigenin; D005189:False Positive Reactions; D006509:Hepatitis B / Q000097:blood / Q000175:diagnosis*; D006514:Hepatitis B Surface Antigens / Q000032:analysis; D006513:Hepatitis B e Antigens / Q000032:analysis; D006515:Hepatitis B virus / Q000235:genetics*; D006526:Hepatitis C / Q000097:blood / Q000175:diagnosis; D006801:Humans; D012680:Sensitivity and Specificity; D014779:Virus Replication / Q000235:genetics
chemical_list: D015342:DNA Probes; D004279:DNA, Viral; D006514:Hepatitis B Surface Antigens; D006513:Hepatitis B e Antigens; D000410:Alanine Transaminase; D004076:Digoxigenin