METHOD FOR HIGH SENSITIVITY DETECTION OF MYD88 MUTATIONS

摘要

A method for increasing sensitivity for detecting minority mutations in MYD88 uses a locked nucleic acid oligo to block amplification of wild-type DNA in DNA isolated from patient FFPE tissue, bone marrow aspirate or peripheral blood samples during PCR while still allowing sequencing and visualization of the PCR product. Further improvement to the sensitivity may be achieved by using a uracil DNA-glycosylase treatment to remove sequence artifacts commonly found in formalin-fixed, paraffin-embedded tissue.

主权项

1. A method for screening and/or monitoring a patient for MYD88 mutations, the method comprising:
isolating DNA from a sampling bone marrow aspirate (BM), fresh peripheral blood (PB), and formalin-fixed, paraffin-embedded (FFPE) tissues sample obtained from the patient suspected of having a condition associated with MYD88 mutations; performing PCR on the extracted DNA to produce amplified DNA while blocking amplification of wild-type DNA; sequencing the amplified DNA in an automated sequencer; analyzing an output of the automated sequencer to identify mutations in the sequence.