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Parasite ; 14(2): 149-54, 2007 Jun.
Article in English | MEDLINE | ID: mdl-17645187

ABSTRACT

We have developed a quantitative PCR assay (LightCycler* using the pair of primers JW58 and JW59 for the detection of the 35-fold repeated B gene of oxoplasma gondii. This real-time PCR, using fluorescence resonance energy transfert (FRET) hybridization probes, allows the quantification of . gondii with several technical requirements not previously described: i) an internal amplification control (co-amplified in a single tube with the same primers), ii) Uracil-N-Glycosylase and iii) a standard curve corresponding to a serial dilution from a calibrated suspension of T. gondii ranging from 40 to 4.106( )parasites in one ml of amniotic fluid (1 to 105( ) . gondii/PCR). In artificial samples, one parasite could be detected if at least three reactions were performed.


Subject(s)
Fluorescence Resonance Energy Transfer/methods , Polymerase Chain Reaction/methods , Toxoplasma/isolation & purification , Toxoplasmosis/diagnosis , Animals , DNA Probes , Gene Amplification , Mice , Nucleic Acid Hybridization , Sensitivity and Specificity , Toxoplasma/genetics
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