Quantitative detection of Toxoplasma gondii DNA in human body fluids by TaqMan polymerase chain reaction.
OBJECTIVE
A new quantitative polymerase chain reaction (real-time PCR) was designed to detect Toxoplasma DNA in human body fluid samples.


METHODS
Real-time fluorescence detection of amplification product formation on the basis of the TaqMan-System was established with Toxoplasma 18S rDNA as a target gene.


RESULTS
The method provides a high sensitivity comparable to conventional nested PCR procedures and generates quantitative data when detecting toxoplasmic DNA in human blood, cerebrospinal or amniotic fluid. Moreover, data were obtained investigating blood samples from an immunocompromised patient with reactivated toxoplasmosis after allogeneic bone marrow transplantation, monitoring the therapeutic effect.


CONCLUSIONS
The potential application of this method to detect Toxoplasma DNA in body fluids and to follow the development of parasitemia under therapy could be demonstrated.
Kupferschmidt
O
O
Robert-Koch-Institut, Parasitologie/Mykologie, Nordufer 20, D-13353 Berlin, Germany. kruegerd@rki.de
Krüger
D
D
Held
T K
TK
Ellerbrok
H
H
Siegert
W
W
Janitschke
K
K
D000818:Animals; D001826:Body Fluids / Q000469:parasitology*; D016026:Bone Marrow Transplantation / Q000009:adverse effects; D016054:DNA, Protozoan / Q000032:analysis*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D019914:Taq Polymerase / Q000378:metabolism; D014122:Toxoplasma / Q000235:genetics / Q000302:isolation & purification*; D014123:Toxoplasmosis / Q000469:parasitology*
D016054:DNA, Protozoan; D019914:Taq Polymerase