Validation d une méthode de Multi-Target-Screening par chromatographie liquide couplée à la spectrométrie de masse en tandem et étude de l utilisation des résultats des criblages toxicologiques dans les services de réanimation et d urgences du CHU de Grenoble

Le criblage toxicologique est une analyse incontournable des laboratoires de toxicologie hospitalière, aussi bien dans le cadre de la prise en charge des intoxications médicamenteuses graves que dans des situations cliniques où l anamnèse reste inconnue et l orientation diagnostique indéfinie. Nous avons dans un premier travail validé une méthode de criblage toxicologique par chromatographie liquide haute performance couplée à la spectrométrie de masse en tandem (CLHP-SM/SM). La détermination des limites d identification a été réalisée pour 117 composés. La variabilité des temps de rétention et des spectres de masse a été évaluée sur un pool de 32 composés. Tous les coefficients de variation analysés étaient inférieurs à 20 %. Ce criblage toxicologique a été validé d un point de vue qualitatif mais une approche semi-quantitative n a pu être validée.Nous avons ensuite évalué de manière prospective sur une période de 6 mois, l utilisation des résultats des criblages toxicologiques dans les services de réanimation et d accueil des urgences du CHU de Grenoble. Quatre-vingt-deux prescriptions ont ainsi été analysées. Le dialogue clinico-biologique a permis de rationaliser les demandes (23 % des prescriptions annulées). Quatre-vingt-dix-sept pourcent des demandes incluses se sont avérées utiles. Les résultats des criblages toxicologiques ont eu une répercussion sur la prise en charge des patients dans 11,1 % des cas. Le dialogue clinico-biologique est indispensable au biologiste médical pour décider de la stratégie des analyses de toxicologie à mettre en place. Ce dialogue est la base d une analyse toxicologique rapide, utile et efficiente.Toxicological screening is an essential analysis in hospital toxicology laboratories, as part of the management of severe drug poisoning in clinical situations where the history is unknown or with undefined diagnostic orientation. First, we validated a method of toxicological screening by high performance liquid chromatography coupled to tandem mass spectrometry (HPLC-MS/MS) .The limits of identification was performed for 117 compounds. Variability in retention time and mass spectra were measured on a pool of 32 compounds. All coefficients of variation were analyzed below 20%. This toxicological screening was validated qualitatively but a semi-quantitative approach could not be validated. Then, we has evaluated prospectively over a period of 6 months, the use of toxicological screening results in intensive care units and emergency of University Hospital of Grenoble. Eighty-two screenings have been analyzed .Clinicobiological explanation has rationalized screenings requirements (23 % of prescriptions were canceled) . Ninety-seven percent of the toxicological screenings were considered useful and had an impact on patient care in 11.1 % of cases. Clinicobiological explanation is essential to help medical biologist to choose the right toxicological analysis strategy to implement. This discussion is the basis for a rapid, helpful and efficient toxicological analysis.GRENOBLE1-BU Médecine pharm. (385162101) / SudocSudocFranceF

Aleukemic congenital leukemia cutis preceding monocytic leukemia with favorable outcome: A case report

Abstract A newborn girl had typical “blueberry muffin” skin lesions, which shows histopathologic features of monocytic leukemia cutis. The systemic leukemia was demonstrated after one month of life. She was treated by chemotherapy, including induction and three consolidation cures, according to the ELAM02 protocol, which led to complete remission. This case report with congenital form of AML5 cutaneous localization, preceding systemic involvement, with a 5‐year follow‐up and positive outcome is remarkable

Agreement in continuous intra-individual robust coefficient of variation between simplified and original flow cytometric gating strategies for quantifying peripheral blood neutrophil myeloperoxidase expression (n = 62).

Abbreviations: RCV = robust coefficient of variation.</p

Comparative diagnostic accuracy of intra-individual robust coefficient of variation for peripheral blood neutrophil myeloperoxidase expression between simplified and original flow cytometric gating strategies (n = 55)^a.

Comparative diagnostic accuracy of intra-individual robust coefficient of variation for peripheral blood neutrophil myeloperoxidase expression between simplified and original flow cytometric gating strategies (n = 55)a.</p

Agreement of binary intra-individual robust coefficient of variation for peripheral blood neutrophil myeloperoxidase expression between simplified and original flow cytometric gating strategies in external validation sample comprising patients with milder level of peripheral blood cytopenia.

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Comparative diagnostic accuracy of intra-individual robust coefficient of variation for peripheral blood neutrophil myeloperoxidase expression between simplified and original flow cytometric gating strategies after excluding two chronic myelomonocytic leukemia cases from the study sample (n = 53).

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Agreement of binary intra-individual robust coefficient of variation for peripheral blood neutrophil myeloperoxidase expression between simplified and original flow cytometric gating strategies after excluding two chronic myelomonocytic leukemia cases from the study sample.

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Deidentified individual participant data.

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Simplified flow cytometric gating strategy for quantifying peripheral blood neutrophil myeloperoxidase expression.

CD45+ cells were first individualized by crossing the singlet gate (A) and CD45 positive gate (B). Population of granulocytes (CD15+ CD14-) was identified (C). Gated cells were selected based on CD16/CD11b double positivity (D) The MPO SSC dot plot (E) was used only to visualize MPO expression of mature neutrophils. The cell population identified was MPO mature neutrophils (red dots). Abbreviations: CD = cluster of differentiation; FSC-A = forward scatter area; FSC-H = forward scatter height; MPO = myeloperoxidase; RCV = robust coefficient of variation; SSC-A = side scatter area; SSC-H = side scatter height.</p

Overview of lasers for FACSCanto-II ^TM flow cytometer.

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