Laboratories

The activity of the Laboratory for Disorders in Hemostasis (Czech abbreviation LPH) is aimed at comprehensive diagnostics of hemostasis: routine and specialized, including translational medicine geared towards disorders in hemostasis. It performs unique examinations of hemostasis in the Czech Republic, such as complete phenotype and genetic screening of patients with a congenital bleeding disease (hemophilia A and B, von Willebrand disease, congenital disorders in other coagulation factors), a congenital thrombotic disease (deficiency of antithrombin,

S protein, C protein) and congenital thrombopathies. Part and parcel of the laboratory is differential diagnostics of TMA (thrombotic microangiopathy), including the atypical hemolytic uremic syndrome (aHUS) test and the thrombotic thrombocytopenic purpura (TTP) test. The quantity of requirements for TMA examination increased considerably in 2020 because the SARS-CoV-2 disease very frequently manifests itself as TMA. The laboratory also focuses on diagnostics of rare acquired disorders in hemostasis (AHA, AWS, iTTP, etc.) and predictive factors of their treatment. It also performs molecular genetic examinations such as the determination of the carrying of hemophilia and prenatal testing of hemophilia and other disorders in hemostasis. When introducing innovative methods of hemostasis examination the laboratory cooperates with cardiological, nephrological, pediatric, neurological, obstetrical, genetic and other centers. It carries out consultation activity in diagnostics of disorders in hemostasis and laboratory diagnostics for the entire Czech Republic. It cooperates in international studies of laboratory monitoring of new anticoagulants and new types of treatment of hemophilia. The laboratory is closely connected with the activity of the Center for Thrombosis and Hemostasis of IHBT. The laboratory is accredited according to the standard ISO:15189. The quality of its activity is monitored through the involvement in external quality control (EQC) of international organizations such as UK NEQAS and ECAT. I tis involved in practical and theoretical training of high-school and university graduates who prepare themselves for specialization exams in hematology and other laboratory fields, including postgraduate (PhD) studies.

The laboratory also acts as the National Reference Laboratory for Disorders in Hemostasis. In this function it organizes annual EQAs of thrombophilia risk factors, supervises over the preparation of material for EQA organizations of SEKK in specialized coagulation methods. On a regular basis, it takes part in the testing of international calibration materials for both routine and highly specialized methods which is organized by the ISTH (International Society for Thrombosis and Hemostasis) and NIBSC (National Institute for Biological Standards and Control). The NRL administers the National Registry of Patients with Congenital Coagulation Disorders comprising data collected since 1980.

The Laboratory of Morphology and Cytochemistry is a laboratory of the Clinical Division that performs routine blood count tests for patients of inpatient centers and the Outpatient Department, including microscopic assessment, and evaluates panoptically stained smears of bone marrow aspirates. Thus, it performs complete cytological diagnostics based on blood count and bone marrow, and for establishing diagnoses it also uses twelve cytochemical methods (see Fig. Bone marrow aspirate smear from a patient with acute myeloid leukemia being stained for the presence of myeloperoxidase with numerous Auer rods in myeloblasts). It offers cytology consultations for most of the Czech Republic. It is one of ten reference laboratories comprising the Group of Expert Centers (Czech abbreviation SEP) for microscopic analysis of peripheral blood smears in the framework of external proficiency testing organized by the SEKK, s. r. o. company Routine diagnostic methods are accredited according to the standard CSN EN ISO:15189 of the CAI company (examination of blood count with an analyzer differential, reticulocytes on an analyzer and microscopic assessment of peripheral blood smear). Blood counts are examined on analyzers made by the SYSMEX CZ company of the XN series, offering other extension parameters (IPF, IG, PLT-F) that can be used in diagnostics of hematological diseases. The efficiency of the device for the preparation of peripheral blood smear and its staining facilitates laboratory work. Digital morphological devices (DM96, DI60) help to evaluate peripheral blood smear, thus contributing an important share to diagnostics of hemato-oncological diseases, chiefly in patients with grave leukopenia and/or neoplasms of the lymphoid line. The laboratory is a year-round training center of high-school and university graduates, including physicians, who prepare themselves for an examination of specialization in hematology

The laboratory performs standard molecular detection of fusion genes in acute myeloid leukemia (AML), including urgent determination of PML/RARα fusion in acute promyelocytic leukemia (APL), plus other fusions of molecular variants of APL. The laboratory performs the capture of clonality of lymphoproliferative diseases of B and T cell origin; in case of chronic lymphatic leukemia (CLL) the laboratory examines the IgHV mutation status (an important prognostic factor). Furthermore, it tests clonal mutations in myeloproliferative neoplasias (MPN): mutations of JAK2 tyrosine kinase, and CALR and MPL gene mutations. It also tests a number of other mutations of prognostic relevance in patients with AML, as well as CLL and MPN. Apart from the above-mentioned disease detection tests, it performs follow-up monitoring of minimal residual disease for AML, APL, CLL and selected cases of MPN. In 2020, the laboratory tested a total of 1,570 blood or bone marrow samples. The core of the laboratory activity is routine diagnostics of the above conditions, and most services are reimbursed from public health insurance. Research activity is connected with the above-mentioned topics. For patients with MPN the laboratory has its own testing panel for detecting other mutations with the help of new generation sequencing.

The Laboratory of Flow Cytometry is a specialized clinical Laboratory for diagnostics and monitoring hemato-oncological diseases (acute leukemia, lymphoproliferative disease, myeloproliferative disease, myelodysplastic syndrome, multiple myeloma) and non-oncological diseases (paroxysmal nocturnal hemoglobinuria, congenital platelet disorders, corpuscular anemia).

Laboratory techniques are validated in conformity with the recommendations of the European Leukemia Net (ELN), the International Clinical Cytometry Society (ICCS), the European Society of Clinical Cell Analysis (ESCCA), and accredited according to the standard CSN EN ISO 15189. Big emphasis is put on the validation of highly sensitive and specific methods for capturing and monitoring measurable residual disease after therapy/transplantation of acute leukemias (AML MRD, B-ALL MRD, T-ALL MRD), lymphoproliferative disorders (B-CLL MRD, MCL MRD, HCL MRD) and multiple myeloma (MM MRD). Annually, the laboratory examines approximately 5,000 patients, and performs some 135,000 procedures for IHBT patients and extra mural consultations for most of the Czech Republic.

The laboratory takes part in research projects and clinical studies, and provides postgradual training of high-school and university graduates in medicine. On a regular basis, it participates in international cycles of external quality assessment (UK NEQAS) and organizes the program of inter-laboratory comparison (ILC) for the Czech and Slovak Republic and the international program of external proficiency testing in the frameworkof the European Society for Clinical Cell Analysis (ESCCA).