Document 0080
 DOCN  M94A0080
 TI    Capillary electrophoresis: detection of hybridization between synthetic
       oligonucleotides and HIV-1 genomic DNA amplified by polymerase-chain
       reaction.
 DT    9412
 AU    Bianchi N; Mischiati C; Feriotto G; Fiorentino D; Di Biase S; Apicella
       N; Gambari R; Biochemistry Institute, Ferrara University, Italy.
 SO    J Virol Methods. 1994 May;47(3):321-9. Unique Identifier : AIDSLINE
       MED/94351052
 AB    The polymerase chain reaction (PCR) is one of the most efficient
       techniques for measuring the viral load of HIV-infected samples.
       Determination of the specificity of PCR products is usually based on
       Southern blotting and hybridization of the amplified DNA to radioactive
       oligonucleotide probes specific for sequences comprised between the PCR
       primers. The recent introduction of capillary electrophoresis (CE) for
       identification of HIV-1 and HTLV-I PCR products appears interesting in
       light of its reproducibility, sensitivity and because it is fast and
       suitable for detection of DNA/DNA and DNA/RNA hybrids. We demonstrate
       that specific hybridization of a HIV-1 oligonucleotide probe to
       single-stranded DNA obtained by unbalanced PCR is detectable by
       capillary electrophoresis. This enabled us the application of a
       one-step, non-radioactive protocol to demonstrate the specificity of
       amplification of HIV-1 genomic sequences by PCR. This procedure is
       simple, reproducible and is suggested as an integral part of automated
       diagnostic systems based on the use of laboratory work stations for DNA
       isolation, preparation of PCR reactions and analysis of PCR products.
 DE    Base Sequence  DNA, Viral/*CHEMISTRY/ISOLATION & PURIF
       Electrophoresis/*METHODS  Genome, Viral  HIV-1/*GENETICS  Molecular
       Sequence Data  Nucleic Acid Hybridization  Oligonucleotides/*CHEMISTRY
       Polymerase Chain Reaction  Support, Non-U.S. Gov't  JOURNAL ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).