title: Preliminary evaluation of one conventional nested and two real-time PCR assays for the detection of Toxoplasma gondii in immunocompromised patients.
abstract: 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.
authors.affiliation: 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.
authors.forename: Thomas
authors.initials: T
authors.lastname: Hierl
authors.affiliation: 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.
authors.forename: Udo
authors.initials: U
authors.lastname: Reischl
authors.affiliation: 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.
authors.forename: Peter
authors.initials: P
authors.lastname: Lang
authors.affiliation: 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.
authors.forename: Holger
authors.initials: H
authors.lastname: Hebart
authors.affiliation: 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.
authors.forename: Maik
authors.initials: M
authors.lastname: Stark
authors.affiliation: 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.
authors.forename: Pierre
authors.initials: P
authors.lastname: Kyme
authors.affiliation: 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.
authors.forename: Ingo B
authors.initials: IB
authors.lastname: Autenrieth
mesh_terms: 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*
chemical_list: D016054:DNA, Protozoan