Evaluation of a Real-time PCR-based assay using the lightcycler system for detection of Toxoplasma gondii bradyzoite genes in blood specimens from patients with toxoplasmic retinochoroiditis

Carlo Contini, Silva Seraceni, Rosario Cultrera, Carlo Incorvaia, Adolfo Sebastiani, Stephan Picot

Research output: Contribution to journalArticle

Abstract

PCR based methods have advantages over traditional methods for the diagnosis of toxoplasmosis, especially when serology fails and clinical symptoms are not evident. However, current PCR-based assays are often labour-intensive and not readily quantifiable and have the potential for contamination due to a requirement for postamplification sample handling. Real-time PCR can address these limitations. We have developed and evaluated a highly sensitive Real-time PCR (Light-cycler, LC-PCR) to detect and quantify Toxoplasma gondii B1 and bradyzoite specific genes (SAG-4, MAG-1) in serum and peripheral blood mononuclear cells (PBMC) specimens, from five immunocompetent subjects with clinically suspected toxoplasmic retinochoroiditis (TRC) or without a suspected T. gondii infection. A standard curve for quantitation of parasitic load was generated using SYBR Green I fluorescent detection. The results were compared with those obtained with a nested PCR (n-PCR). In TRC patients, both PCR methods confirmed ophtalmoscopy and fluorangiographic findings. Among the TRC patients, the use of LC-PCR was more sensitive than n-PCR for detection and quantification of either B1 gene (P

Original languageEnglish
Pages (from-to)275-283
Number of pages9
JournalInternational Journal for Parasitology
Volume35
Issue number3
DOIs
Publication statusPublished - Mar 2005

Keywords

  • Ocular toxoplasmosis
  • PCR
  • Real-time PCR
  • Toxoplasma gondii
  • Toxoplasmic chorioretinitis

ASJC Scopus subject areas

  • Parasitology
  • Infectious Diseases

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