Development and Prospective Validation of a Standardized Flow Cytometric Assay of Peripheral Blood Neutrophil Myeloperoxidase Expression for Ruling Out Myelodysplastic Syndromes. (MPO-MDS-Develp)
|The safety and scientific validity of this study is the responsibility of the study sponsor and investigators. Listing a study does not mean it has been evaluated by the U.S. Federal Government. Read our disclaimer for details.|
|ClinicalTrials.gov Identifier: NCT04399018|
Recruitment Status : Active, not recruiting
First Posted : May 22, 2020
Last Update Posted : January 9, 2023
Myelodysplastic syndromes (MDS) comprise a heterogeneous group of clonal bone marrow neoplasms that predominate in the elderly, with a median age at diagnosis of 70 years. The diagnosis of MDS relies on peripheral blood cytopenia and morphologic dysplasia for one or more hematopoietic cell lineage. Cytopenia is evidenced with hemogram while dysplasia requires bone marrow aspirate, which is an invasive procedure .
Considering the low prevalence of disease among subjects referred for suspected MDS, many patients are exposed to unnecessary bone marrow aspiration-related discomfort and harms. Therefore, an objective assay based on a peripheral blood sample that accurately discriminates MDS from other cytopenia etiologies is highly desirable.
We have previously developed and refined a flow cytometric analysis protocol for quantifying neutrophil MPO expression in peripheral blood at three university-affiliated hospitals (i.e., Clermont-Ferrand, Saint-Etienne, and Grenoble) (Raskovalova et al, Hematologica 2019). We found that the robust coefficient of variation (RCV, computed as the robust standard deviation divided by the median) within an individual subject was the best parameter in discriminating patients with versus without MDS.
Although promising, flow cytometric analysis of neutrophil MPO expression in peripheral blood is technically complex, time consuming, and not standardized. Hence, its performance requires specific expertise and the results show substantial variability. A single ready-to-use tube with lyophilized antibodies would have the potential to standardize the measurement of neutrophil MPO expression in peripheral blood across laboratories, with results available within 30-60 min in routine practice.
In this study, the investigators hypothesize that a standardized and semi-automatic flow cytometric assay of neutrophil MPO expression in peripheral blood could accurately rule out MDS and obviate the need for bone marrow aspiration and biopsy, with sensitivity and negative predictive value estimates approaching 100%.
In this observational diagnostic accuracy study, burden will be null for recruited patients. No specific intervention is assigned to participants. All diagnostic testing, procedures, and medication ordering are performed at the discretion of attending physicians. A test result will have no impact on patient management. .Compliance with current guidelines disseminated by the French Haute Autorité de Santé (HAS) will be advocated for the diagnostic work-up of patients with suspected MDS. No follow-up visits are planned in this cross-sectional study.
|Condition or disease||Intervention/treatment|
|Myelodysplastic Syndromes||Other: Diagnostic Test: Flow cytometry analysis of neutrophil myeloperoxidase expression|
|Study Type :||Observational|
|Actual Enrollment :||103 participants|
|Official Title:||Development and Prospective Validation of a Standardized Flow Cytometric Assay of Peripheral Blood Neutrophil Myeloperoxidase Expression for Ruling Out Myelodysplastic Syndromes.|
|Actual Study Start Date :||July 27, 2020|
|Estimated Primary Completion Date :||July 2024|
|Estimated Study Completion Date :||December 2024|
- Other: Diagnostic Test: Flow cytometry analysis of neutrophil myeloperoxidase expression
Flow cytometry analysis of neutrophil myeloperoxidase expression in peripheral blood samples will be performed within 24 h of MDS diagnostic evaluation and blinded to the reference standard. Peripheral blood samples will be collected in 5 ml (EDTA) anticoagulant plastic tubes and processed within 24 h maximum of collection.
Blood sample will be stained according to the manufacturers' recommendations with a lyophilized cocktail ("LyotubeTM ready-to-use", BD Bioscience).
At least 10,000 neutrophils will be acquired on a 3-laser, 8-color BD FACSCanto-II TM flow cytometer (BD Biosciences, San José, CA).
Each marker will be expressed as median, geometric and arithmetic mean, regular and robust coefficient of variation.
- Reference diagnosis of MDS or CMML established by bone marrow examination [ Time Frame: Baseline ]The primary outcome is the reference diagnosis of MDS or CMML established by bone marrow examination (with Perls staining) by two independent experienced hematopathologists blinded to the index test results. Disagreements will be solved by a third hematopathologist. Bone marrow aspirate will be repeated within 4 to 6 months for patients with idiopathic cytopenia of uncertain significance (ICUS) or non-conclusive bone marrow examination.
- Intra-laboratory coefficient of variation for intra-individual RCV, computed as the standard deviation multiplied by 100 and divided by the mean (intra- and inter-assay precision) [ Time Frame: Baseline ]
- Relative change from baseline, expressed in percentages for intra-individual RCV [ Time Frame: Baseline ]
- Inter-laboratory coefficient of variation for intra-individual RCV [ Time Frame: Baseline ]
- Negative predictive value point estimates (along with 95% confidence interval) of neutrophil myeloperoxidase expression in peripheral blood for the diagnosis of MDS or CMML [ Time Frame: Baseline ]
To learn more about this study, you or your doctor may contact the study research staff using the contact information provided by the sponsor.
Please refer to this study by its ClinicalTrials.gov identifier (NCT number): NCT04399018
|Chu Grenoble Alpes|
|Grenoble, France, 38043|
|Study Chair:||Tatiana Raskovalova, MD||Centre Hospitalier Universitaire Grenoble Alpes|
|Principal Investigator:||Richard Veyrat-Masson, MD||Centre Hospitalier Universitaire Clermont-Ferrand|
|Principal Investigator:||Carmen Aanei, MD||Centre Hospitalier Universitaire Saint-Étienne|