title: Multicentric comparative assessment of the bio-evolution Toxoplasma gondii detection kit with eight laboratory-developed PCR assays for molecular diagnosis of congenital toxoplasmosis.
abstract: The detection of Toxoplasma gondii in amniotic fluid is an essential tool for the prenatal diagnosis of congenital toxoplasmosis and is currently essentially based on the use of PCR. Although some consensus is emerging, this molecular diagnosis suffers from a lack of standardization and an extreme diversity of laboratory-developed methods. Commercial kits for the detection of T. gondii by PCR were recently developed and offer certain advantages; however, they must be assessed in comparison with optimized reference PCR assays. The present multicentric study aimed to compare the performances of the Bio-Evolution T. gondii detection kit and laboratory-developed PCR assays set up in eight proficient centers in France. The study compared 157 amniotic fluid samples and found concordances of 99% and 100% using 76 T. gondii-infected samples and 81 uninfected samples, respectively. Moreover, taking into account the classification of the European Research Network on Congenital Toxoplasmosis, the overall diagnostic sensitivity of all assays was identical and calculated to be 86% (54/63); specificity was 100% for all assays. Finally, the relative quantification results were in good agreement between the kit and the laboratory-developed assays. The good performances of this commercial kit are probably in part linked to the use of a number of good practices: detection in multiplicate, amplification of the repetitive DNA target rep529, and the use of an internal control for the detection of PCR inhibitors. The only drawbacks noted at the time of the study were the absence of uracil-N-glycosylase and small defects in the reliability of the production of different reagents.
authors.affiliation: Institut de Parasitologie et de Pathologie Tropicale, Université de Strasbourg, Laboratoire de Parasitologie et Mycologie Médicale, Hôpitaux Universitaires de Strasbourg, Strasbourg, France Pôle Biologie Moléculaire du Centre National de Référence de la Toxoplasmose, Montpellier, France.
authors.forename: Denis
authors.initials: D
authors.lastname: Filisetti
authors.affiliation: Centre Hospitalier Universitaire (CHU) of Montpellier/University Montpellier I, Laboratoire de Parasitologie-Mycologie, UMR CNRS 5290/IRD 224/UM, Montpellier, France Pôle Biologie Moléculaire du Centre National de Référence de la Toxoplasmose, Montpellier, France.
authors.forename: Yvon
authors.initials: Y
authors.lastname: Sterkers
authors.affiliation: CHU A. Michallon/University Joseph Fourier Grenoble I, Laboratoire de Parasitologie-Mycologie, Département des Agents Infectieux, Grenoble, France Pôle Biologie Moléculaire du Centre National de Référence de la Toxoplasmose, Montpellier, France.
authors.forename: Marie-Pierre
authors.initials: MP
authors.lastname: Brenier-Pinchart
authors.affiliation: CHU of Toulouse, Service de Parasitologie-Mycologie, Toulouse, France Pôle Biologie Moléculaire du Centre National de Référence de la Toxoplasmose, Montpellier, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Cassaing
authors.affiliation: CHU of Dijon, Laboratoire de Parasitologie-Mycologie, Faculty of Medicine, Dijon, France Pôle Biologie Moléculaire du Centre National de Référence de la Toxoplasmose, Montpellier, France.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Dalle
authors.affiliation: CHU of Lille/University Nord de France (UDSL), Service de Parasitologie-Mycologie, Faculty of Medicine, Institut Pasteur de Lille, Lille, France Pôle Biologie Moléculaire du Centre National de Référence de la Toxoplasmose, Montpellier, France.
authors.forename: Laurence
authors.initials: L
authors.lastname: Delhaes
authors.affiliation: CHU A. Michallon/University Joseph Fourier Grenoble I, Laboratoire de Parasitologie-Mycologie, Département des Agents Infectieux, Grenoble, France Pôle Biologie Moléculaire du Centre National de Référence de la Toxoplasmose, Montpellier, France.
authors.forename: Hervé
authors.initials: H
authors.lastname: Pelloux
authors.affiliation: AP-HP, Groupe Hospitalier Pitié-Salpêtrière, Laboratoire de Parasitologie-Mycologie, Paris, France Pôle Biologie Moléculaire du Centre National de Référence de la Toxoplasmose, Montpellier, France.
authors.forename: Fériel
authors.initials: F
authors.lastname: Touafek
authors.affiliation: Centre Hospitalier Universitaire (CHU) of Montpellier/University Montpellier I, Laboratoire de Parasitologie-Mycologie, UMR CNRS 5290/IRD 224/UM, Montpellier, France University Montpellier 1, Faculty of Pharmacy, Laboratoire de Biophysique & Bioanalyse, Montpellier, France Pôle Biologie Moléculaire du Centre National de Référence de la Toxoplasmose, Montpellier, France.
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Varlet-Marie
authors.affiliation: Université Paris-Descartes, AP-HP, Groupe Hospitalier Cochin, Laboratoire de Parasitologie-Mycologie, Paris, France Pôle Biologie Moléculaire du Centre National de Référence de la Toxoplasmose, Montpellier, France.
authors.forename: Hélène
authors.initials: H
authors.lastname: Yera
authors.affiliation: Institut de Parasitologie et de Pathologie Tropicale, Université de Strasbourg, Laboratoire de Parasitologie et Mycologie Médicale, Hôpitaux Universitaires de Strasbourg, Strasbourg, France.
authors.forename: Ermano
authors.initials: E
authors.lastname: Candolfi
authors.affiliation: Centre Hospitalier Universitaire (CHU) of Montpellier/University Montpellier I, Laboratoire de Parasitologie-Mycologie, UMR CNRS 5290/IRD 224/UM, Montpellier, France Pôle Biologie Moléculaire du Centre National de Référence de la Toxoplasmose, Montpellier, France p-bastien@chu-montpellier.fr.
authors.forename: Patrick
authors.initials: P
authors.lastname: Bastien
mesh_terms: D000653:Amniotic Fluid / Q000469:parasitology; D015331:Cohort Studies; D005260:Female; D006801:Humans; D059021:Laboratory Proficiency Testing; D016133:Polymerase Chain Reaction / Q000379:methods / Q000592:standards; D011247:Pregnancy; D011933:Reagent Kits, Diagnostic / Q000592:standards; D015203:Reproducibility of Results; D012189:Retrospective Studies; D012680:Sensitivity and Specificity; D014122:Toxoplasma / Q000235:genetics*; D014125:Toxoplasmosis, Congenital / Q000175:diagnosis* / Q000469:parasitology*
chemical_list: D011933:Reagent Kits, Diagnostic