Characterization and multicentric validation of a common standard for Toxoplasma gondii detection using nucleic acid amplification assays
| Autor: | Yvon Sterkers, Laurence Delhaes, Sophie Cassaing, Hervé Pelloux, Denis Filisetti, Emmanuelle Varlet-Marie, Feriel Touafek, Marie-Pierre Brenier-Pinchart, Hélène Yera, Patrick Bastien, Frédéric Dalle |
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| Přispěvatelé: | Laboratoire de Parasitologie-Mycologie (CHU de Montpellier), Pôle Biologie-Pathologie [CHRU Montpellier], Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier)-Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier), Université de Montpellier (UM), Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier), Laboratoire de parasitologie-mycologie, CHU Grenoble, Service de Parasitologie et Mycologie, CHU Toulouse [Toulouse]-Institut Fédératif de Biologie (IFB) - Hôpital Purpan, Hôpital Purpan [Toulouse], CHU Toulouse [Toulouse]-CHU Toulouse [Toulouse]-Hôpital Purpan [Toulouse], CHU Toulouse [Toulouse], Laboratoire de parasitologie mycologie (CHU de Dijon), Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon), FLAveur, VIsion et Comportement du consommateur (FLAVIC), Institut National de la Recherche Agronomique (INRA)-Etablissement National d'Enseignement Supérieur Agronomique de Dijon (ENESAD)-Université de Bourgogne (UB), Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP), Service de Parasitologie-Mycologie [CHRU LIlle], Institut de Microbiologie [CHRU Lille], Pôle de Biologie Pathologie Génétique [CHU Lille], Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Pôle de Biologie Pathologie Génétique [CHU Lille], Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille), Laboratoire de parasitologie et mycologie médicale [CHU Strasbourg], CHU Strasbourg, Service de parasitologie - mycologie [CHU Pitié-Salpétrière], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-CHU Pitié-Salpêtrière [AP-HP], Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Service de parasitologie-mycologie [CHU Cochin], Hôpital Cochin [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), We acknowledge the financial support of the 'RSI Assurance Maladie Professions Libérales—Provinces, C.A.M.P.L.P.' for buying the LightCycler 480 (Roche) real-time PCR equipment in Montpellier as well as that of the Institut de Veille Sanitaire (InVS)., We thank Florence Robert-Gangneux (CHU of Rennes) and Jean Menotti (Hôpital Saint-Louis, Paris) for fruitful discussions. We are grateful to Sylvie Matern and Rachel Huber (Strasbourg), Filomena Naji and Michèle Wauquier (Lille), Sylvie Douzou, Ghislaine Serres, and Bounleth Sanichanh (Montpellier), Catherine Barois (Grenoble), Anne Lamy, Raphaelle Laurenceau, Sandrine Chalmeton, Elodie Duthu, Séverine Gisquet, and Catherine Paris (Toulouse), and Cédric Rondot and Yacine Garaoun (Paris-Cochin) for technical assistance. We are indebted to Nathalie Van Langendonck (Academic Hospital of Tours) for providing AF samples. We are also grateful to Sebastien Besteiro (UMR CNRS 5235/University Montpellier 2, 'DIMNP') for providing the in vitro cultivated parasites., Pôle biologie Pathologie [CHRU Montpellier], Service de Parasitologie-Mycologie [Toulouse], Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-CHU Toulouse [Toulouse]-Hôpital Purpan [Toulouse], Agroécologie [Dijon], Institut National de la Recherche Agronomique (INRA)-Université de Bourgogne (UB)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Université Bourgogne Franche-Comté [COMUE] (UBFC), Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-CHU Pitié-Salpêtrière [APHP], Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-CHU Cochin [AP-HP] |
| Rok vydání: | 2014 |
| Předmět: |
Microbiology (medical)
MESH: Reference Standards MESH: Molecular Diagnostic Techniques/methods MESH: Parasite Load/standards [SDV]Life Sciences [q-bio] Toxoplasma gondii diagnostic Parasitic loads Parasite Load MESH: Nucleic Acid Amplification Techniques/standards MESH: Toxoplasma/isolation & purification medicine Molecular diagnostic techniques Humans National level Reference standards MESH: Parasite Load/methods standardization MESH: Humans biology MESH: Nucleic Acid Amplification Techniques/methods Nucleic acid amplification technique MESH: Toxoplasmosis/diagnosis MESH: Molecular Diagnostic Techniques/standards Reference Standards biology.organism_classification medicine.disease Virology Toxoplasmosis quantification 3. Good health MESH: France Molecular Diagnostic Techniques Immunology Nucleic acid MESH: Toxoplasma/genetics Parasitology France Nucleic Acid Amplification Techniques Toxoplasma |
| Zdroj: | Journal of Clinical Microbiology Journal of Clinical Microbiology, American Society for Microbiology, 2014, 52 (11), pp.3952-3959. ⟨10.1128/jcm.01906-14⟩ Journal of Clinical Microbiology, American Society for Microbiology, 2014, 52 (11), pp.3952-3959. ⟨10.1128/JCM.01906-14⟩ |
| ISSN: | 1098-660X 0095-1137 |
| Popis: | The molecular diagnosis of toxoplasmosis essentially relies upon laboratory-developed methods and suffers from lack of standardization, hence the large diversity of performances between laboratories. Moreover, quantifications of parasitic loads differ among centers, a fact which prevents the possible prediction of the severity of this disease as a function of parasitic loads. The objectives of this multicentric study performed in eight proficient laboratories of the Molecular Biology Pole of the French National Reference Center for Toxoplasmosis (NRC-T) were (i) to assess the suitability of a lyophilized preparation of Toxoplasma gondii as a common standard for use in this PCR-based molecular diagnosis and (ii) to make this standard available to the community. High-quality written procedures were used for the production and qualification of this standard. Three independent batches of this standard, containing concentrations ranging from 10 4 to 0.01 T. gondii genome equivalents per PCR, were first assessed: the linear dynamic range was ≥6 log, the intra-assay coefficients of variation (CV) from a sample containing 10 T. gondii organisms per PCR were 0.3% to 0.42%, and the interassay CV over a 2-week period was 0.76% to 1.47%. A further assessment in eight diagnostic centers showed that the standard is stable, robust, and reliable. These lyophilized standards can easily be produced at a larger scale when needed and can be made widely available at the national level. To our knowledge, this is the first quality control assessment of a common standard which is usable both for self-evaluation in laboratories and for accurate quantification of parasitic loads in T. gondii prenatal infections. |
| Databáze: | OpenAIRE |
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