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Gewählte Publikation:

Publikationstyp: Zeitschriftenaufsatz
Dokumenttyp: Originalarbeit

Jahr: 1999

AutorInnen: Leutenegger, CM; Klein, D; Hofmann-Lehmann, R; Mislin, C; Hummel, U; Böni, J; Boretti, F; Guenzburg, WH; Lutz, H

Titel: Rapid feline immunodeficiency virus provirus quantitation by polymerase chain reaction using the TaqMan fluorogenic real-time detection system.

Quelle: J Virol Methods. 1999; 78(1-2):105-116



Autor/innen der Vetmeduni Vienna:

Günzburg Walter
Klein Dieter

Beteiligte Vetmed-Organisationseinheiten
Institut für Virologie


Zugehörige(s) Projekt(e): Virus-Wirt Interaktionen


Abstract:
An improved quantitative polymerase chain reaction (qPCR) method based on a combination of real-time detection and the 5"-3" nuclease activity of the Taq DNA polymerase was developed to quantify the provirus load of feline immunodeficiency virus (FIV), a lentivirus of veterinary importance and an animal model for AIDS research. Two fluorogenic probes were designed to detect FIV provirus in genomic DNA of peripheral lymphocytes and tissues infected with different FIV subtypes. The most sensitive assay can detect one copy of FIV provirus. The assay showed excellent precision within-run and between-runs. Comparison of the TaqMan system with a conventional seminested PCR assay revealed a comparable detection limit and good correlation. Furthermore the design of the two probes allowed the detection of various FIV isolates of clade A and B.

Keywords Pubmed: Animals
Cats
DNA Primers
DNA, Viral/analysis
Feline Acquired Immunodeficiency Syndrome/virology*
Fluorescent Dyes
Immunodeficiency Virus, Feline/genetics
Immunodeficiency Virus, Feline/isolation & purification*
Oligonucleotide Probes
Polymerase Chain Reaction/methods*
Proviruses/isolation & purification*
Sensitivity and Specificity
Taq Polymerase/metabolism*


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