Abstract：
: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.
journal_name
J Virol Methodsjournal_title
Journal of virological methodsauthors
Bianchi N,Mischiati C,Feriotto G,Fiorentino D,Di Biase S,Apicella N,Gambari Rdoi
10.1016/0166-0934(94)90028-0subject
Has Abstractpub_date
1994-05-01 00:00:00pages
321-9issue
3eissn
0166-0934issn
1879-0984pii
0166-0934(94)90028-0journal_volume
47pub_type
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