Forschung

Laboratory for Chimerism and MRD

Molecular biologic analyses have become an integral part of diagnostic and follow-up investigation of live threatening malignant and non-malignant diseases, thus completing cytogenetic and immunological investigations.

Our laboratory supports the evaluation of treatment follow-up mainly in patients after allogeneic blood stem cell transplantation. After enrichment of special cellular fractions and cell subpopulations we apply qualitative and quantitative PCR procedures, fragment analysis and nucleotide sequencing.

The focus of our laboratory is the analysis of chimerism after allogeneic stem cell transplantation and the analysis of minimal residual disease (MRD) in patients with acute lymphoblastic leukemia. Both procedures are certified according to DIN EN ISO 15189(Certificate) . The laboratory is certified according to DIN EN ISO 9001:2008, it is member of the Euro-chimerism consortium and member of ESLOH EuroMRD-ALL.

    Chimerism analysis:

    The aim of transplantation is the eradication the recipient’s haematopoietic system and its permanently replacement by the haematopoietic system of the donor. Complete chimerism is established when all haematopoietic cells originate from the donor. In malignant diseases like leukaemia complete chimerism is associated with a lower risk of relapse.

      • By means of chimerism analysis characteristic microsatellite profiles can be detected in samples of recipient and donor.
      • The investigation of subcellular fractions (enriched by MACS-Beads) increases the limit of detection.
      • Required sample material:
        A reference sample from donor and recipient is required once before transplantation. Sample size for follow-up depends on the number of subcellular fractions (2 ml EDTA/requested investigation). The results are available on average 1-2 working days after receipt of the sample.
      • If you have further questions, please contact Prof. Bader or Dr. Kreyenberg.

        Detection of Minimal Residual Disease in Acute Lymphoblastic Leukemia:

        ALL is the most frequent malignant disease in children. The success of treatment is dependent on permanent elimination of leukemic cells below microscopic detection limit.

        The standard method for the investigation of BCR-ABL negative ALL is the detection of patient’s leukaemia specific somatically rearranged immunoglobuline (Ig-) and T-cell receptor (TCR) genes. The procedure is divided into 2 steps:

          • 1. Characterization of junctional sequences and design of complementary PCR-primer
          • 2. MRD detection in follow-up samples by means of quantitative PCR
          • Required sample material:
            One initial diagnostic sample of the leukemia (as reference). 5 ml EDTA-BM or
            10 ml EDTA-PB is necessary. The results of the follow-up analysis (log-reduction compared with reference sample) are available on average 2-4 working days after receipt of the sample.

            Further procedures:

            • Identity analysis (for example of cultured cells, cell lines or tumour samples)
            • Determination of feto-maternal microchimerism by means of Real-Time PCR
            • Vß-spectratyping to control immune reconstitution
            • Quantitative analysis of WT1 expression
            • Sequencing analysis

              Contact

              Head
              Dr. rer. nat. Hermann Kreyenberg

              Mo-Fr 9-17
              Laboratoy
              Phone: +49 (0) 69 6301-6449
              Fax: +49 (0) 69 6301-83937

              Scientific advice and information
              Phone: +49 (0) 69 6301-7542 (Prof. Bader)
              Phone: +49 (0) 69 6301-83926
              (Dr. Kreyenberg)

              Theodor-Stern-Kai 7
              60590 Frankfurt
              Building 32 A, Second floor