Adapting a conventional PCR assay for Toxoplasma gondii detection to real-time quantitative PCR including a competitive internal control.
Parasite
; 14(2): 149-54, 2007 Jun.
Article
in En
| 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.
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Collection:
01-internacional
Health context:
2_ODS3
/
3_ND
Database:
MEDLINE
Main subject:
Toxoplasma
/
Toxoplasmosis
/
Polymerase Chain Reaction
/
Fluorescence Resonance Energy Transfer
Type of study:
Diagnostic_studies
Limits:
Animals
Language:
En
Journal:
Parasite
Year:
2007
Document type:
Article