Competitive polymerase chain reaction for quantitating feline immunodeficiency virus load in infected cat tissues

Mauro Pistello, Stefano Menzo, Massimo Giorgi, Luigi Da Prato, Giancarlo Cammarota, Massimo Clementi, Mauro Bendinelli

Research output: Contribution to journalArticle

Abstract

To quantitate FIV provirus copy numbers present in tissue of infected cats, we have applied a competitive polymerase chain reaction (cPCR) recently described for HIV. The method consists in coamplifying a fixed amount of the DNA to be examined with graded copy numbers of a DNA competitor incorporating a short deletion and bearing the same primer recognition sequences. These conditions ensure almost identical thermodynamic and amplification efficiency for both template species but permit a prompt recognition of the two amplification products by gel electrophoresis. Since the amounts of the two amplicons are dependent on relative initial template concentrations, the number of FIV genomes in the samples can be calculated by densitometric analysis of the electrophoretics bands. After validation, the method has been applied to study the provirus loads in the tissues of cats infected with the Pisa-M2 isolate of FIV.

Original languageEnglish
Pages (from-to)229-234
Number of pages6
JournalMolecular and Cellular Probes
Volume8
Issue number3
DOIs
Publication statusPublished - Jun 1994

Keywords

  • quantitative PCR, feline immunodeficiency virus, competitive PCR, feline-AIDS

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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