Preliminary evaluation of one conventional nested and two real-time PCR assays for the detection of Toxoplasma gondii in immunocompromised patients.
Toxoplasma reactivation is a serious complication in patients receiving allogenic stem cell transplantation. Real-time PCR assays allow a rapid diagnosis of toxoplasma infection; however, no comparative data are available on the performance of real-time PCR protocols under routine conditions. Therefore, the aim of this study was to amplify Toxoplasma gondii DNA from routine samples of allogenic stem cell recipients using two real-time PCR assays on a LightCycler, and using conventional nested PCR. Conventional nested PCR revealed T. gondii DNA in 16 samples. Only 12 of the 16 samples yielded a positive result in both real-time PCRs. The accuracy of the conventional PCR results was demonstrated by direct sequencing. Amplification and detection of the amplicon was completed in only 1 h using the real-time PCR assays. Thus, real-time PCR substantially accelerates the detection of T. gondii DNA in the majority of positive specimens; however, conventional nested PCR is required for detection of T. gondii DNA in some samples.
Hierl
Thomas
T
Departments of Medical Microbiology and Hospital Hygiene1, Pediatrics3 and Hematology and Oncology, Medical Clinic4, University of Tübingen, Germany 2Department of Medical Microbiology, University of Regensburg, Germany.
Reischl
Udo
U
Departments of Medical Microbiology and Hospital Hygiene1, Pediatrics3 and Hematology and Oncology, Medical Clinic4, University of Tübingen, Germany 2Department of Medical Microbiology, University of Regensburg, Germany.
Lang
Peter
P
Departments of Medical Microbiology and Hospital Hygiene1, Pediatrics3 and Hematology and Oncology, Medical Clinic4, University of Tübingen, Germany 2Department of Medical Microbiology, University of Regensburg, Germany.
Hebart
Holger
H
Departments of Medical Microbiology and Hospital Hygiene1, Pediatrics3 and Hematology and Oncology, Medical Clinic4, University of Tübingen, Germany 2Department of Medical Microbiology, University of Regensburg, Germany.
Stark
Maik
M
Departments of Medical Microbiology and Hospital Hygiene1, Pediatrics3 and Hematology and Oncology, Medical Clinic4, University of Tübingen, Germany 2Department of Medical Microbiology, University of Regensburg, Germany.
Kyme
Pierre
P
Departments of Medical Microbiology and Hospital Hygiene1, Pediatrics3 and Hematology and Oncology, Medical Clinic4, University of Tübingen, Germany 2Department of Medical Microbiology, University of Regensburg, Germany.
Autenrieth
Ingo B
IB
Departments of Medical Microbiology and Hospital Hygiene1, Pediatrics3 and Hematology and Oncology, Medical Clinic4, University of Tübingen, Germany 2Department of Medical Microbiology, University of Regensburg, Germany.
D000818:Animals; D016054:DNA, Protozoan / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification*; D005260:Female; D006801:Humans; D016867:Immunocompromised Host; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D014122:Toxoplasma / Q000235:genetics / Q000302:isolation & purification*; D014123:Toxoplasmosis / Q000469:parasitology*
D016054:DNA, Protozoan