Automated determination of amplified PCR products: application to HCV viremia detection and quantification.
A simple and reliable automated method was developed for the detection of amplified products after PCR, which provides an alternative to the time-consuming Southern blotting and hybridization procedure. For the determination and quantification of hepatitis C viremia, the digoxigenin labelling process was applied during the PCR of the amplicons, followed by an inverse hybridization assay performed with biotinylated probes. The detection of the PCR amplified products could be processed as simply as an ELISA, or by means of an automated analyser.
Clossais Besnard
N
N
Laboratorie de Bactériologie-Virologie, Centre Hospitalo-Universitaire, Pontchaillou, Rennes, France.
Minjolle
S
S
Gicquel
J
J
Colimon
R
R
André
P M
PM
D001483:Base Sequence; D015139:Blotting, Southern; D017931:DNA Primers / Q000235:genetics; D015342:DNA Probes / Q000235:genetics; D004076:Digoxigenin; D004797:Enzyme-Linked Immunosorbent Assay; D005069:Evaluation Studies as Topic; D005784:Gene Amplification; D006526:Hepatitis C / Q000175:diagnosis* / Q000821:virology; D006801:Humans; D015336:Molecular Probe Techniques / Q000706:statistics & numerical data; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D012367:RNA, Viral / Q000097:blood / Q000235:genetics; D012680:Sensitivity and Specificity; D014766:Viremia / Q000175:diagnosis* / Q000821:virology
D017931:DNA Primers; D015342:DNA Probes; D012367:RNA, Viral; D004076:Digoxigenin