Laboratory of PCR Diagnostics of Leukemia

The lab is a molecular genetic laboratory that performs molecular detection of fusion genes in acute myeloid leukemia (AML), detection of clonality in lymphoproliferative diseases of B and T series - in chronic lymphocytic leukemia (CLL; in addition, it determines the mutational status of IgHV genes and the presence of mutations in the TP53 gene), or other chronic and acute lymphoproliferative diseases. In addition, the laboratory investigates clonal mutations in Ph-negative myeloproliferative neoplasia (MPN) - JAK2V617F tyrosine kinase mutation and CALR gene mutations. In addition to these investigations, when disease is detected, the laboratory performs follow-up molecular monitoring of minimal residual disease (MRD) in both AML and CLL, and in selected cases of MPN.

As part of the diagnosis of acute myeloid leukemia, the laboratory screens each patient for the PML/RARα, AML1/ETO and CBFβ/MYH11 fusion genes and for the presence of an internal tandem duplication of the FLT3 gene. Based on the cytogenetic findings, various MLL gene fusions with different breakpoints and possibly other fusion genes can be investigated. The laboratory also has the option to investigate alternative fusion genes that may rarely occur in atypical forms of acute promyelocytic leukemia that do not carry the commonly investigated PML/RARα fusion gene but may have a PLZF/RARα, NPM1/RARα, NuMA/RARα, FIP1L1/RARα, STA5b/RARα, PRKAR1A/RARα, BCOR/RARα, or OBFC2A/RARα fusion. Molecular characterization of a specific fusion gene is then used to monitor MRD using real-time quantitative RT-PCR (RQ-PCR).

In the context of lymphoproliferative diseases in CLL, the laboratory investigates clonal rearrangements of IgHV genes (in patients with CLL or some lymphomas) and evaluates their mutational status. It also investigates TP53 gene mutations by direct cDNA sequencing.

In T-cell lymphoproliferative disorders, it determines clonal rearrangements of the δ and γ chains of the T-cell receptor (TCR) genes. By appointment, the laboratory performs MRD testing by RQ-PCR using allele-specific primers in patients with previously demonstrated clonal IgHV or TCR rearrangements.

In MPN, the laboratory routinely determines the presence of the JAK2V617F mutation using RQ-PCR with LNA-modified probes. Alternative JAK2 gene mutations in exon 12 can also be investigated in selected patients with polyglobulia. In patients with thrombocytosis or myelofibrosis, the laboratory detects CALR gene length mutations by fragmentation analysis. In selected patients with MPN (e.g., those who do not carry known mutations in the JAK2, CALR or MPL genes, or those with a family history of MPNs), next-generation sequencing can also be performed to detect less common mutations.