Mentype® Chimera® PCR Amplification Kit

The gold standard for STR analysis

Features

High-throughput cost-effective solution for pre- and post-transplantation application
10 % better chance to find highly informative, polymorphic discrimination markers
Monitoring of a wide chimerism range with superior sensitivity of ≥ 1.4 %
Registered CE-IVD product according to EU 2017/746 (IVDR)

Mentype^® Chimera^® PCR Amplification Kit is a multiplex assay for qualitative genotyping and semi-quantitative chimerism monitoring after allogeneic hematopoietic stem cell transplantation. The assay employs capillary electrophoresis technology with minimal DNA input and demonstrates a superior detection limit compared to other STR-PCR methods - capable of identifying recipient fractions as low as 1.4%.

Biomarkers

12 tetra- / pentanucleotide STR loci, including 6 STR markers validated by European Consortium of Chimerism Analysis, and 1 sex-specific INDEL:

Amelogenin (Xp22.1-22.3, Yp11.2), D2S1360 (2p24-p22), D3S1744 (3p24), D4S2366(4p16-15.2), D5S2500 (5q11.2), D6S474 (6q21-22), D7S1517 (7q31.33), D8S1132 (8q23.1), D10S2325 (10p12), D12S391 (12p13.2)

Product Specifications

Panel
12 STRs + Amelogenin

Reactions
1 multiplex PCR reaction per sample

PCR controls
2 included (PC, NTC)

Sample input
0.125 - 2 ng gDNA from peripheral blood

Sensitivity
≥ 1.4 %

Turnaround time
~ 4.5 h after DNA extraction

Detection
Qualitative genotyping, Semiquantitative chimerism detection

To be used with
Standard thermal cycler + Thermo Fisher Genetic Analyzer

Data analysis
ChimerisMonitor IVD, GeneMapper^TM ID-X + specific templates

Accessory
Matrix Standard BT5 multi

Scientific Background

Allogeneic hematopoietic stem cell transplantation (HSCT) is a treatment option for patients with non-malignant and malignant hematological diseases, such as leukemia. Chimerism analysis is used to determine the mixture of donor and recipient hematopoietic cells in allo-HSCT recipients to detect early signs of graft rejection. Human peripheral venous blood is used for genotyping and monitoring. Depending on the success of transplantation, different forms of hematopoietic chimerism (complete, mixed or loss) can develop. Different approaches are used for chimerism analysis, including fluorescence in situ hybridization (FISH), restriction fragment length polymorphism (RFLP), blood count analysis and PCR-based methods. Currently, PCR-based amplification of short tandem repeat (STR) polymorphisms is the golden standard for chimerism analysis. To detect early signs of graft rejection, chimerism analysis should be done at regular intervals and shortly after the allogeneic HSCT.

Product References

Carbonell, D. et al. FLT3-ITD Expression as a Potential Biomarker for the Assessment of Treatment Response in Patients with Acute Myeloid Leukemia. Cancers 14. https://doi.org/10.3390/cancers14164006 (2022)
Iranzo-Tatay, C. et al. Genome-wide DNA methylation profiling in anorexia nervosa discordant identical twins. Translational Psychiatry 12, https://doi.org/10.1038/s41398-021-01776-y (2022)
Benítez, L. et al. The Contribution of QF-PCR and Pathology Studies in the Diagnosis of Diandric Triploidy/Partial Mole. Diagnostics 11. https://doi.org/10.3390/diagnostics11101811 (2021)
Ruhnke, L. et al. Long-Term Mixed Chimerism After Ex Vivo/In Vivo T Cell-Depleted Allogeneic Hematopoietic Cell Transplantation in Patients With Myeloid Neoplasms. Front Oncol 11. https://doi.org/10.3389/fonc.2021.776946 (2021)
Handisurya, A. et al. Antigen-Specific Immunoadsorption With the Glycosorb® ABO Immunoadsorption System as a Novel Treatment Modality in Pure Red Cell Aplasia Following Major and Bidirectional ABO-Incompatible Allogeneic Hematopoietic Stem Cell Transplantation. Front Med 7, https://doi.org/10.3389/fmed.2020.585628 (2020)
Navarro-Bailón, A et al. Short Tandem Repeats (STRs) as Biomarkers for the Quantitative Follow-Up of Chimerism after Stem Cell Transplantation: Methodological Considerations and Clinical Application. Genes 11, 993. https://doi.org/10.3390/genes11090993 (2020)
Ventura-Aguiar, P. et al. Late Onset Graft Plasmacytoma-Like PTLD Presenting as Acute Hyperglycemia in a Kidney-Pancreas Transplant Recipient. Case Reports in Nephrology 2019. https://doi.org/10.1155/2019/2818074 (2019)
Martín, I. et al. Therapy-related acute myeloid leukemia developing 14 years after allogeneic hematopoietic stem cell transplantation, from a persistent R882H-DNMT3A mutated clone of patient origin. Experimental and Molecular Pathology 105, 139-143. https://doi.org/10.1016/j.yexmp.2018.07.002 (2018)
Rautenberg, C. et al. Wilms' Tumor 1 Gene Expression Using a Standardized European LeukemiaNet-Certified Assay Compared to Other Methods for Detection of Minimal Residual Disease in Myelodysplastic Syndrome and Acute Myelogenous Leukemia after Allogeneic Blood Stem Cell Transplantation. Biol Blood Marrow Transplant 24, 2337-2343. https://doi.org/10.1016/j.bbmt.2018.05.011 (2018)
Ahci, M. et al. Clinical Utility of Quantitative PCR for Chimerism and Engraftment Monitoring after Allogeneic Stem Cell Transplantation for Hematologic Malignancies. Biol Blood Marrow Transplant 23, 1658-1668. https://doi.org/10.1016/j.bbmt.2017.05.031 (2017)
Clemente, I. et al. Relevance of Chimerism Analysis After Allogeneic Stem Cell Transplantation. Transplant Proc 49, 890-892. https://doi.org/10.1016/j.transproceed.2017.01.065 (2017)
Rasche, L. et al. Allogeneic Hematopoietic Cell Transplantation in Multiple Myeloma: Focus on Longitudinal Assessment of Donor Chimerism, Extramedullary Disease, and High-Risk Cytogenetic Features. Biol Blood Marrow Transplant 22, 1988-1996. https://doi.org/10.1016/j.bbmt.2016.08.024 (2016)
Stahl, T. et al. Digital PCR Panel for Sensitive Hematopoietic Chimerism Quantification after Allogeneic Stem Cell Transplantation. Int. J. Mol. Sci. 17. https://doi.org/10.3390/ijms17091515 (2016)
Stahl, T. et al. Digital PCR to assess hematopoietic chimerism after allogeneic stem cell transplantation. Experimental Hematology 43, 462-468. https://doi.org/10.1016/j.exphem.2015.02.006 (2015)
Platzbecker, U. et al. Azacitidine for treatment of imminent relapse in MDS or AML patients after allogeneic HSCT: results of the RELAZA trial. Leukemia 26, 381-389. https://doi.org/10.1038/leu.2011.234 (2012)
Bornhäuser, M. et al. Prophylactic transfer of BCR-ABL–, PR1-, and WT1-reactive donor T cells after T cell–depleted allogeneic hematopoietic cell transplantation in patients with chronic myeloid leukemia. Blood 117, 7174-7184. https://doi.org/10.1182/blood-2010-09-308569 (2011)
Bornhäuser, M. et al. Monitoring of donor chimerism in sorted CD34+ peripheral blood cells allows the sensitive detection of imminent relapse after allogeneic stem cell transplantation. Haematologica 94, 1613-1617, https://doi.org/10.3324%2Fhaematol.2009.007765 (2009)