Method, reagent and kit for detecting minimal residual disease

A kit and reagent technology, applied in the field of cancer diagnosis, can solve problems such as hindering low-frequency malignant cell detection

Active Publication Date: 2020-06-26
ERASMUS UNIV MEDICAL CENT ROTTERDAM ERASMUS MC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it should be noted that a high frequency of normal regenerative cells can interfere with the detection of low frequencies of malignant cells in blood, bone marrow, and other bodily fluids (e.g., cerebrospinal fluid) during and after treatment and after hematopoietic stem cell transplantation

Method used

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  • Method, reagent and kit for detecting minimal residual disease
  • Method, reagent and kit for detecting minimal residual disease
  • Method, reagent and kit for detecting minimal residual disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0096] Example 1. Antibody panel and diagnostic method for MRD detection in BCP-ALL patients

[0097] Markers used to identify total B cells and B cell precursors in bone marrow

[0098] List of relevant identification markers : CD19, CD45

[0099] How to use them : The use of CD19 markers to preset gates is necessary to identify pure B cell populations. To focus on normal B cell precursors (BCP), CD45 negative or weakly positive can be used to distinguish CD45 positive mature B cells from BCP. In the case of CD19 treatment, CD22 can be used instead of CD19. These markers can also be used in combination with side light scattering (SSC) or forward light scattering (FSC) or both FSC and SSC to identify peripheral blood or bone marrow or other types of samples (e.g. bone marrow, tissue biopsy, spinal fluid) B cells in. It is worth noting that other markers used to distinguish between normal BCP cells and BCP-ALL cells (such as CD10, CD20, CD38, and CD34) (see below) can also help...

Embodiment 2

[0114] Example 2. Antibody panel and diagnostic method for MRD detection in CLL patients

[0115] Markers used to identify total B cells in peripheral blood and bone marrow:

[0116] List of identification markers :CD19, CD3 (exclude markers)

[0117] How to use them : Using this combination of markers to preset gates is necessary to identify pure B cell populations and remove T cell / B cell doublets. These markers can also be used in combination with side light scattering (SSC) or forward light scattering (FSC) or both FSC and SSC to identify peripheral blood or bone marrow or other types of samples (e.g., tissue biopsy, spinal fluid) B cells. For finer gating to better enrich CLL cells, both CD5 and CD27 can be used. Markers used to distinguish between normal B cells and CLL cells:

[0118] List of markers and the most common phenotypic aberrations :

[0119] CD27 : Positive on CLL cells and a small part of normal B cells

[0120] CD5 : Positive on CLL cells and a small part o...

Embodiment 3

[0129] Example 3. Antibody panel and diagnostic method for MRD detection in MM / PCD patients

[0130] Markers used to identify total plasma cells in bone marrow:

[0131] List of identification markers : CD38, CD138 and CD229

[0132] How to use them: Any combination of the three markers works at any fluorescent dye position; any combination of two of the three markers can also be used or even one of the three markers alone can be used in some cases (not all). It is preferably combined in the following order: 1) CD138 / CD38 / CD229; 2) CD138 / CD38, 3) CD138 / CD229; 4) CD38 / CD229; 5) CD138; 6) CD38); 7) CD229. It should be noted that any of these markers, alone and in combination, can also be used in combination with side light scattering (SSC) or forward light scattering (FSC) or both FSC and SSC to identify bone marrow or other types of samples (e.g., peripheral blood, Tissue biopsy, spinal fluid) in plasma cells.

[0133] Markers used to distinguish between normal and clonal / malignan...

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Abstract

The invention relates to a method, a reagent and a kit for detecting minimal residual disease. The present invention relates to the field of minimal residual disease (MRD) diagnostics, which is increasingly used to evaluate the efficacy of treatment in patients with hematological malignancies, such as B-cell precursor acute lymphoblastic leukemia (BCP-ALL), B-cell chronic lymphocytic Cellular Leukemia (B‑CLL) and Multiple Myeloma (MM). The unique reagent composition offered features carefully selected and thoroughly tested antibody combinations that achieve at least 10 for ≥8 color flow cytometry staining and for 10 and 12 color flow cytometry ‑4 even as low as 10 ‑5 sensitivity. The invention also provides diagnostic kits and methods for detecting MRD.

Description

[0001] This application is a divisional application of the Chinese patent application with the application number 201380037624.4. The original application is a PCT international application PCT / NL2013 / 050420 filed on June 14, 2013, which entered the Chinese national phase on January 14, 2015. Technical field [0002] The present invention relates to the field of cancer diagnosis, and more particularly to means and methods for monitoring disease development during and after treatment or for detecting micro-disseminated diseases. Suppressive cell therapy or cytotoxic agent therapy induces remission in most patients with lymphoid malignancies. However, many of these patients will relapse. Obviously, although they have achieved the so-called complete remission based on cell morphological standards, the existing suppressive cell therapy or cytotoxic agent treatment regimens cannot kill all malignant cells in these relapsed patients. Because the detection limit of cell morphology tech...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/574G01N15/14
CPCG01N33/57426G01N15/14G01N33/57484G01N2015/1006A61K38/164A61K38/1716A61K39/025A61K39/09
Inventor 雅各布斯·约翰内斯·玛丽亚·万东恩何塞·阿尔贝托·奥尔方德马托斯克瑞亚埃韦尔朱安·亚历杭德罗·弗洛雷斯蒙特罗茱莉亚·玛丽亚·阿尔梅达帕拉文森特·亨利库斯·约翰内斯·万德费尔登塞巴斯蒂安·博埃特赫尔朗托尼·威廉·朗格拉克埃斯特尔·迈斯特日科娃托马斯·什切潘斯基马蒂亚斯·里特根保罗·约根·蒙泰罗达席尔瓦卢西奥
Owner ERASMUS UNIV MEDICAL CENT ROTTERDAM ERASMUS MC
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