Competitive polymerase chain reaction using an internal standard: application to the quantitation of viral DNA

Détails

ID Serval
serval:BIB_CFE8F64EB5C8
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Competitive polymerase chain reaction using an internal standard: application to the quantitation of viral DNA
Périodique
Journal of Virological Methods
Auteur(s)
Telenti  A., Imboden  P., Germann  D.
ISSN
0166-0934 (Print)
Statut éditorial
Publié
Date de publication
09/1992
Volume
39
Numéro
3
Pages
259-68
Notes
Journal Article
Research Support, Non-U.S. Gov't --- Old month value: Sep
Résumé
A general strategy for the construction of an internal standard for the polymerase chain reaction (PCR) is described together with its application in the evaluation of clinical samples. This internal standard is a plasmid containing a modified target sequence that is co-amplified with the native target using the same set of primers. The co-amplification reaction will generate two fragments of different size that are readily separated without the need for restriction enzyme digestion. Thereafter, they are detected and quantitated by hybridization to the same probe. Detection of HIV proviral DNA was chosen as a model for this competitive PCR. The assay proved to be a sensitive tool for the detection of PCR inhibitors and allowed quantitation of HIV with a 20-30% variation coefficient. Despite limitations that appear inherent to the amplification process, internal standards appear to be useful tools for quantitative analysis by PCR.
Mots-clé
Base Sequence Binding, Competitive Cloning, Molecular DNA, Viral/*analysis HIV Infections/diagnosis Humans Molecular Sequence Data Polymerase Chain Reaction/*methods/*standards
Pubmed
Web of science
Création de la notice
25/01/2008 14:45
Dernière modification de la notice
20/08/2019 15:50
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