Digoxygenin-labelled DNA-probe: a rapid non-radioactive method for hepatitis B virus DNA detection in serum.
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.
Buti
M
M
Department of Internal Medicine, Hospital Valle Hebrón, Universidad Autónoma, Barcelona, Spain.
Jardi
R
R
Rodriguez-Frias
F
F
Arranz
J A
JA
Casacuberta
J M
JM
Sansegundo
B
B
Puigdomenech
P
P
Esteban
R
R
Guardia
J
J
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
D015342:DNA Probes; D004279:DNA, Viral; D006514:Hepatitis B Surface Antigens; D006513:Hepatitis B e Antigens; D000410:Alanine Transaminase; D004076:Digoxigenin