title: Detection of NOTCH1 c.7541_7542delCT mutation in chronic lymphocytic leukemia using conventional and real-time polymerase chain reaction.
abstract: To evaluate real-time polymerase chain reaction (PCR) assay system for detection of NOTCH1 c.7541_754delCT mutation in chronic lymphocytic leukemia (CLL) patients.
  
  
  METHODS
  A total of 325 CLL patients were included in the study. Screening for NOTCH1 c.7544_7545delCT was performed using conventional PCR-based amplification refractory mutation system (ARMS) method. All 33 samples harboring c.7544_7545delCT allele and 5 negative cases as control were submitted to real-time PCR.
  
  
  RESULTS
  Specificity and sensitivity of two PCR techniques were comparable. NOTCH1 c.7544_7545delCT mutation was found by ARMS in 10.1% of CLL patients, which is consistent with the data of other studies. However, the results of ARMS PCR in a minority of cases (2.15%) were doubtful and required reinvestigation. Real-time PCR, being less time-consuming, showed advantage in the assessment of the amplification's specificity (using the melting curve analysis). It also allows the quantitative assessment of NOTCH1-mutated clone.
  
  
  CONCLUSIONS
  NOTCH1 c.7544_7545delCT mutation resulting in removal of the C-terminal PEST domain, deregulation of NOTCH1-dependent signaling pathways, has negative influence on prognosis of CLL and efficiency of therapy with anti-CD20 monoclonal antibodies. Real-time PCR allows the fast and reliable detection of c.7544_7545delCT mutation and can be used for the screening of this molecular lesion in CLL patients.
authors.affiliation: SI "National Research Center for Radiation Medicine of the National Academy of Medical Sciences of Ukraine", Kyiv 04050, Ukraine.
authors.forename: N I
authors.initials: NI
authors.lastname: Bilous
authors.affiliation: SI "National Research Center for Radiation Medicine of the National Academy of Medical Sciences of Ukraine", Kyiv 04050, Ukraine.
authors.forename: I V
authors.initials: IV
authors.lastname: Abramenko
authors.affiliation: SI "National Research Center for Radiation Medicine of the National Academy of Medical Sciences of Ukraine", Kyiv 04050, Ukraine.
authors.forename: A A
authors.initials: AA
authors.lastname: Chumak
authors.affiliation: SI "National Research Center for Radiation Medicine of the National Academy of Medical Sciences of Ukraine", Kyiv 04050, Ukraine.
authors.forename: I S
authors.initials: IS
authors.lastname: Dyagil
authors.affiliation: SI "National Research Center for Radiation Medicine of the National Academy of Medical Sciences of Ukraine", Kyiv 04050, Ukraine.
authors.forename: Z V
authors.initials: ZV
authors.lastname: Martіna
mesh_terms: D001483:Base Sequence; D016368:Frameshift Mutation; D006801:Humans; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000175:diagnosis / Q000235:genetics*; D011379:Prognosis; D060888:Real-Time Polymerase Chain Reaction; D051881:Receptor, Notch1 / Q000235:genetics*; D017384:Sequence Deletion
chemical_list: C497761:NOTCH1 protein, human; D051881:Receptor, Notch1