Detection of notch1 c.7544_7545deICT mutation in chronic lymphocytic leukemia using conventional and real-time polymerase chain reaction

• Published in: Experimental Oncology
• Date: 2016
• Main Authors: Bilous, N.I., Abramenko, I.V., Chumak, A.A., Dyagil, I.S., Martina, Z.V.
• Format: Article
• Language: English
• Published: Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України 2016
• Subjects: Original contributions
• Online Access: https://nasplib.isofts.kiev.ua/handle/123456789/138001
• Journal Title: Digital Library of Periodicals of National Academy of Sciences of Ukraine
• Cite this: Detection of notch1 c.7544_7545deICT mutation in chronic lymphocytic leukemia using conventional and real-time polymerase chain reaction / N.I. Bilous, I.V. Abramenko, A.A. Chumak, I.S. Dyagil, Z.V. Martina // Experimental Oncology. — 2016 — Т. 38, № 2. — С. 112–116. — Бібліогр.: 28 назв. — англ.

Aim: To evaluate real-time polymerase chain reaction (PCR) assay system for detection of NOTCH1 c.7541_754delCT mutation in chronic lymphocytic leukemia (CLL) patients. Material and 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. Conclusion: 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.