PCR Diagnostics Laboratory for Leukemias

The workplace is incorporated into the Complement of laboratories IHBT. It continuously upholds independent and impartial assessment of the professional competence of the Complement of laboratories in accordance with the ČSN EN ISO 15189 standard, whose application is verified by the Czech Institute for Accreditation. Examination methods within the scope of accreditation are defined in the appendix of the Accreditation Certificate in the current version, which is available on the website IHBT

The laboratory is a molecular genetic laboratory that performs molecular detection of fusion genes in acute myeloid leukemia (AML), clonality detection in lymphoproliferative diseases of B and T series – in chronic lymphocytic leukemia (CLL; in which it additionally determines the mutational status of IgHV genes and the presence of mutations in the TP53 gene), or other chronic and acute lymphoproliferative diseases. It further examines clonal mutations in Ph myeloproliferative neoplasms (MPN) – the mutation of tyrosine kinase JAK2V617F and mutations of the CALR gene. In addition to the aforementioned examinations for disease detection, the laboratory also performs subsequent molecular monitoring of minimal residual disease (MRD) in AML and CLL, and in selected cases of MPN.

In determining the diagnosis of acute myeloid leukemia, the laboratory examines fusion genes in each patient PML/RARα, AML1/ETO and CBFβ/MYH11 and the presence of internal tandem duplication of the FLT3 gene. Based on the cytogenetic finding, it is possible to examine various fusions of the MLL gene with different breakpoints, or possibly other fusion genes. The laboratory further has the possibility to examine alternative fusion genes that may rarely occur in atypical forms of acute promyelocytic leukemia that do not carry the commonly examined fusion gene PML/RARα, but may have fusion PLZF/RARα, NPM1/RARα, NuMA/RARα, FIP1L1/RARα, STA5b/RARα, PRKAR1A/RARα, BCOR/RARα, or OBFC2A/RARα. The molecular characterization of a specific fusion gene is subsequently used for MRD monitoring using quantitative RT-PCR in real time (RQ-PCR).

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

In T – cell lymphoproliferative diseases, it determines clonal rearrangements of δ and γ chain genes of T – cell receptor (TCR). By agreement, the laboratory performs MRD examination using RQ-PCR with allele-specific primers in patients with previously demonstrated clonal rearrangement of IgHV or TCR.

In myeloproliferative diseases, the laboratory routinely determines the presence of JAK2V617F mutation using the RQ-PCR method with LNA-modified probes. In selected patients with polycythemia, alternative JAK2 gene mutations in exon 12 can also be examined. In patients with thrombocytosis or myelofibrosis, the laboratory detects length mutations of the CALR gene by fragment analysis. In selected patients with MPN (for example, those who do not carry known mutations of the JAK2, CALR or MPL genes, or in people with a family history), it is possible to perform examination using next-generation sequencing to detect less frequent mutations.

Employees are bound by confidentiality pursuant to the Act on health services, 372/2011 Coll.

Trust and high data security is one of the basic priorities in the activities of the Complement of laboratories IHBT. Established procedures proceed in accordance with the Personal Data Processing Act 110/2019 Coll., and Regulation (EU) 2016/679 of the European Parliament and of the Council of 27 April 2016 on the protection of natural persons with regard to the processing of personal data and on the free movement of such data and repealing Directive 95/46/EC. More information is available on the website IHBT.