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NRL for DNA Diagnostics

The NRL for DNA diagnostics is integrated in the Complement of the laboratories of the IHBT. Since 2002, it has continuously advocated independent and impartial assessment of professional competence according to ČSN EN ISO/IEC 17025 Conformity assessment - General requirements for the competence of testing and calibration laboratories, the application of which is verified by the Czech Institute for Accreditation. The current version of the accreditation certificate is available on the IHBT website.

The NRL for DNA diagnostics comprises three separate departments: the Molecular Genetics Department, the HLA Department and the Cell Chimerism Department, which are also accredited by the CIA, o.p.s. within the Complement of Laboratories according to the ČSN EN ISO 15189 Standard for Medical Laboratories - Quality and Competence Requirements. The main professional focus of the NRL is molecular genetic diagnostics of selected haemato-oncological diseases, accurate determination of HLA genotypes and determination of cellular chimerism.

In an accredited testing laboratory we perform testing of diagnostic kits, provide control samples (DNA) with defined characteristics ensuring continuity with international standards and verify results for other laboratories. Individual departments are also involved in research activities and serve as training facilities. Our services include the organisation of specialised training courses, seminars and the provision of expert advice.

The departments organise external quality control (inter-laboratory comparison tests ) every year.

For the needs or at the customer's request, the NRL for DNA diagnostics can use the granted "flexible scope of accreditation" to introduce new or modified methods or testing procedures (new instrumentation) within the already granted scope of accreditation, under the conditions of maintaining the principle of testing. The current list of methods and procedures within the granted flexible scope of accreditation is given here: List of activities within the flexible scope of accreditation.

Documents for download:

 

HLA Department

Department: Kateřinská 521/19, 120 00 Prague 2

Head of department. Milena Vraná

Deputy Head: Mgr. Mgr. Radek Plachý

Currently, we perform genotyping of HLA class I and II loci: HLA A, B, C, DRB1, DQB1, -DQA1, DPB1 at the level of determination of allelic groups or single alleles, and determination of KIR genes. If requested by the sponsors, the offer can be extended to other classical and non-classical HLA genes.

Polymerase chain reaction (PCR) and massively parallel sequencing (NGS) based methods are used for testing.

The control materials are also used as samples for the quality control provided by MPZ (Detection of HLA alleles associated with diseases).

Tests performed within the scope of accreditation:

  • Determination of gene sequence variants associated with haematological and immunological diseases and haematopoietic cell transplantation programme by PCR with gel electrophoresis (NRL_05_SOP_14_02 procedure A)
  • Determination of gene sequence variants related to haematological and immunological diseases and haematopoietic cell transplantation programme by Real-time PCR (NRL_05_SOP_14_02 procedure C)
  • Determination of sequence variants of genes related to haematological and immunological diseases and the haematopoietic stem cell transplant programme by NGS-MPS (NRL_05_SOP_14_02 procedure D)

 

Department of Molecular Genetics

Department: U Nemocnice 2094/1, 128 00 Prague 2

Head of the department: doc.Mgr. Kateřina Machová Poláková, Ph.D.

Deputy Head: Mgr. Hana Žižková, Ph.D.

The focus of the testing laboratory is to determine the presence and type of BCR::ABL1 fusion gene rearrangement, to determine the level of BCR::ABL1 transcript and to detect mutations in the BCR::ABL1 kinase domain that cause resistance to treatment with tyrosine kinase inhibitors.

We are also preparing control materials, which we use for regularly organized MITs, for the BCR::ABL1 fusion gene testing.

Tests performed within the scope of accreditation:

  • Determination of the type of BCR::ABL1 fusion gene rearrangement by multiplex RT PCR (NRL_03_SOP_14_02)
  • Determination of BCR::ABL1 transcript levels by Real-time RT PCR (NRL_04_SOP_14_02/procedure A)
  • Determination of mutations in the kinase domain of BCR::ABL1 by Sanger direct sequencing (NRL_04_SOP_14_02/procedure B)
  • Determination of a conversion factor for expressing BCR::ABL1 transcript level results internationally (NRL_04_SOP_14_02/procedure C)

 

Department of Cell Chimerism

Department: Kateřinská 521/19, 120 00 Prague 2

Head of the department. Mgr. Lucie Stefflová

Deputy Head: RNDr. Monika Leontovyčová

The genotyping of sequence polymorphisms is used to identify the cell population (procedure A). If more than one cell population is present in the sample, the quantification of the individual cell populations follows (procedure B). For identification and quantification, methods such as PCR and fragment analysis of short tandem repeats (STRs), and real-time PCR of short insertions and deletions (indels) are utilized.

These procedures can also be applied for cell line authentication, which some scientific journals recommend prior to the publication of a peer-reviewed article. The department also offers testing for the genetic match of different biological samples.

Tests performed within the scope of accreditation:

  • Genotyping of sequence polymorphisms by PCR with gel electrophoresis (NRL_01_SOP_14_02/ procedure A)

  • Determination of the quantitative representation of genotypes in a sample by PCR with gel electrophoresis of sequence polymorphism (NRL_01_SOP_14_02, procedure B)

  • Determination of genotypes of sequence polymorphisms by Real-time PCR (NRL_07_SOP_14_02, procedure A)

  • Determination of the quantitative representation of genotypes in a sample by Real-time PCR of sequence polymorphisms (NRL_07_SOP_14_02, procedure B)

 

The Department of Cell Chimerism annually organizes the external proficiency testing.

The staff is bound by confidentiality according to the Health Services Act, 372/2011 Coll. Confidence and high data security are among the basic priorities within the activities of the Complement of Laboratories of the IHBT. The established procedures are carried out 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 physical 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 IHBT website.