Detection of hepatitis B virus DNA in human serum samples: use of digoxigenin-labeled oligonucleotides as modified primers for the polymerase chain reaction.
Nonradioactive techniques have been used for the direct detection of hepatitis B virus DNA in human serum samples. A comparison of two different systems using digoxigenin-labeled DNA probes is presented. Furthermore, oligonucleotides containing one molecule of the hapten digoxigenin at the 5'-end were prepared and used as primers for the polymerase chain reaction. Amplified DNA can be directly analyzed with anti-digoxigenin Fab fragments labeled with alkaline phosphatase and chemiluminescent substrates.
Escarceller
M
M
Department of Molecular Genetics, CID-CSIC, Barcelona, Spain.
Rodriguez-Frias
F
F
Jardi
R
R
San Segundo
B
B
Eritja
R
R
D000469:Alkaline Phosphatase; D001483:Base Sequence; D015342:DNA Probes / Q000138:chemical synthesis; D004279:DNA, Viral / Q000097:blood*; D004076:Digoxigenin; D005784:Gene Amplification; D006509:Hepatitis B / Q000097:blood; D006515:Hepatitis B virus / Q000235:genetics*; D006801:Humans; D007128:Immunoglobulin Fragments; D008163:Luminescent Measurements; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D009841:Oligonucleotides / Q000138:chemical synthesis; D016133:Polymerase Chain Reaction / Q000379:methods*
D015342:DNA Probes; D004279:DNA, Viral; D007128:Immunoglobulin Fragments; D015345:Oligonucleotide Probes; D009841:Oligonucleotides; D000469:Alkaline Phosphatase; D004076:Digoxigenin