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Development and Prospective Validation of a Standardized Flow Cytometric Assay of Peripheral Blood Neutrophil Myeloperoxidase Expression for Ruling Out Myelodysplastic Syndromes. (MPO-MDS-Develp)

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ClinicalTrials.gov Identifier: NCT04399018
Recruitment Status : Active, not recruiting
First Posted : May 22, 2020
Last Update Posted : January 9, 2023
Information provided by (Responsible Party):
University Hospital, Grenoble

Brief Summary:

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

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Study Type : Observational
Actual Enrollment : 103 participants
Observational Model: Cohort
Time Perspective: Cross-Sectional
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

Resource links provided by the National Library of Medicine

Intervention Details:
  • 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.

Primary Outcome Measures :
  1. 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.

Secondary Outcome Measures :
  1. 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 ]
  2. Relative change from baseline, expressed in percentages for intra-individual RCV [ Time Frame: Baseline ]
  3. Inter-laboratory coefficient of variation for intra-individual RCV [ Time Frame: Baseline ]
  4. 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 ]

Information from the National Library of Medicine

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Ages Eligible for Study:   18 Years to 100 Years   (Adult, Older Adult)
Sexes Eligible for Study:   All
Accepts Healthy Volunteers:   No
Sampling Method:   Probability Sample
Study Population
Eligible participants are unselected consecutive adults referred for suspected MDS. Suspicion of MDS is based on medical history and peripheral blood cytopenia. To be eligible, patients will be required to meet all five inclusion criteria and none of the exclusion criteria.

Inclusion Criteria:

  • Age at enrollment ≥18 years
  • Clinical suspicion of MDS
  • Indication for bone marrow examination
  • ≥1 peripheral blood cytopenia defined by hemoglobin concentration <12 g/dL for female and <13g/dL for male patients, platelet count <150 x109/L, absolute neutrophil count <1.8 x109/L
  • Inpatient and outpatient care patients

Exclusion Criteria:

  • Refusal to participate
  • History of or active documented MDS
  • Enrollment in intensive or critical care unit
  • Incarcerated or individuals protected by French regulation (Article L1121.5 and following, Code de la Santé Publique)
  • Not affiliated with social security system
  • Previous enrollment in the study.

Information from the National Library of Medicine

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

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Chu Grenoble Alpes
Grenoble, France, 38043
Sponsors and Collaborators
University Hospital, Grenoble
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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
Publications automatically indexed to this study by ClinicalTrials.gov Identifier (NCT Number):
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Responsible Party: University Hospital, Grenoble
ClinicalTrials.gov Identifier: NCT04399018    
Other Study ID Numbers: 38RC19.425
2019-A03341-56 ( Other Identifier: ID RCB )
First Posted: May 22, 2020    Key Record Dates
Last Update Posted: January 9, 2023
Last Verified: January 2023
Individual Participant Data (IPD) Sharing Statement:
Plan to Share IPD: Yes
Plan Description: The principal investigators will respond directly to data requests by providing a de-identified data set. Individual participant data that underlie the results reported in the published articles (i.e., main text, tables, figures, and appendices) will be supplied.
Supporting Materials: Study Protocol
Statistical Analysis Plan (SAP)
Time Frame: No later than 3 years after final acceptance of the primary study paper.
Access Criteria:

De-identified data will be available for individual participant data meta-analysis purpose.

Researchers should submit a methodologically sound proposal that complies with the Preferred Reporting Items for Systematic Review and Meta-Analysis Protocols (PRISMA-P) 2015 statement. Proposals should be directed to TRaskovalova@chu-grenoble.fr. Data requestors will need to sign a data access agreement.

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Studies a U.S. FDA-regulated Drug Product: No
Studies a U.S. FDA-regulated Device Product: No
Keywords provided by University Hospital, Grenoble:
Diagnostic accuracy study
Flow cytometry analysis
Peripheral blood sample
Additional relevant MeSH terms:
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Myelodysplastic Syndromes
Pathologic Processes
Bone Marrow Diseases
Hematologic Diseases
Precancerous Conditions