diagnostics of chromosomal translocations of the MLL-gene for the diagnosis of acute leukaemia
INNOVECTIS Gesellschaft für Innovations-Dienstleistungen mbH
Molecular diagnostics of chromosomal translocations of the MLL-gene are of high importance for the identification and therapy monitoring of acute leukaemia due to the causally interrelation of translocations and the development of the acute syndromes (lymphatic or myelocytic).
About 1,300 people additionally fall ill with acute leukaemia every year. The chances of healing are rather low because of frequent relapses. Therefore an individual care with good diagnostic opportunities is of high interest.
State of the art methods like FISH and several PCR-methods (like RT, inverse, pan-handle PCR) show several disadvantages, like low resolution (approx. 10-4) and therefore missing identification of fusion genes; inevitable knowledge of the translocation partner genes involved; quality of the used genetic information carriers (like RNA) and more. Comprehensive, meaningful analyses require high time and effort (months to years) and are not suitable for routine use.
In order to guarantee a routine diagnosis and effective therapy care of high-risk-leukaemia-patients a reliable, extensive and fast diagnostic procedure is required. The presented innovation covers these advantages. The in-vitro technique identifies translocations with genomic DNA via LDI-PCR (Long Distance Inverse-PCR). In addition, the method is suitable for MRD-examinations (Minimal Residual Disease) in the case of relapses.
Translocations, deletions or duplications are identified as chromosomal abberations of the human MLL-gene.
Unknown translocations/partner genes, duplications and deletions can be identified. In addition, “MLL-internal duplications” (cause leukaemia as well) are recognized.
The required samples are derived from bone marrow-, blood- or biopsy samples from which the genomic DNA is isolated.
The limit of detection is one cancer cell per 100,000 to 1,000,000 normal cells.
The genomic DNA is religated to circles, amplified by LDI-PCR and analysed by gel electrophoresis/sequence determination. Different pairs of oligonucleotides are applied for amplification.
Due to an early identification of a MLL-translocation, treatments with special protocols for chemotherapy can be performed.
In the case of relapses the procedure can be applied for MRD-examinations to guarantee an early action and a valuable monitoring of the examinations. Specific DNA-sequences are raised for each patient and used as a marker.
The method can be used as a matter of routine; therefore it is available as a kit.
The method has already been applied at the Frankfurt University Hospital.
A German patent application is filed.An international application based on the PCT regulations is filed.
Owner of patent
Johann Wolfgang Goethe University Frankfurt
Licensing or assignment of the technology is possible as well as a cooperation for further development of the invention.