Related Experiment Video
Updated: Mar 20, 2026
Author Spotlight: Advancing the Detection of Low-Frequency Mutations in Cancer Tissues
Published on: August 23, 2024
ABL gene kinase domain mutation scanning by denaturing high performance liquid chromatography sequencing method
Yücel Erbilgin, Suzin Çatal, Ahmet Emre Eşkazan
1Department of Genetics, Institute of Experimental Medicine, İstanbul University, İstanbul, Turkey Phone: +90 212 414 20 00-33312
Objective:
Despite the efficacy of the BCR-ABL tyrosine kinase inhibitor imatinib, the development of resistance against imatinib has been observed. The most important mechanisms known to cause resistance are point mutations in the ABL tyrosine kinase and the ATP domain. This study describes the use of denaturing high performance liquid chromatography (dHPLC) as a method to screen for mutations of the ABL gene.
Methods:
We used the dHPLC based assay for the screening of ABL point mutations. Forty chronic myeloid leukemia (CML) patients who showed resistance to imatinib were screened in parallel by dHPLC and direct sequencing.
Results:
Nine of the 40 patients (23%) had mutations.
Conclusion:
dHPLC can be a useful method for pre-screening. Analyzing the mutations and monitoring (high-risk) patients can improve their prognosis and survival rate. dHPLC can potentially become a valuable tool for regular testing of patients in the future.
Insights
Denaturing high-performance liquid chromatography (dHPLC) effectively screens for ABL gene mutations in imatinib-resistant chronic myeloid leukemia (CML) patients. This method aids in identifying resistance mechanisms, potentially improving patient prognosis and survival rates.
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- Imatinib is a crucial BCR-ABL tyrosine kinase inhibitor for chronic myeloid leukemia (CML).
- Resistance to imatinib often arises from point mutations in the ABL kinase or ATP-binding domain.
- Accurate and efficient mutation detection is vital for managing imatinib resistance.
Purpose of the Study:
- To evaluate denaturing high-performance liquid chromatography (dHPLC) as a screening method for ABL gene mutations.
- To assess the prevalence of ABL mutations in CML patients resistant to imatinib.
Main Methods:
- A dHPLC-based assay was employed to screen for ABL point mutations.
- Forty imatinib-resistant CML patients were analyzed using both dHPLC and direct sequencing for parallel comparison.
Main Results:
- Mutations were identified in nine out of the 40 (23%) CML patients screened.
- dHPLC demonstrated its capability to detect these resistance-associated mutations.
Conclusions:
- dHPLC serves as a valuable pre-screening tool for ABL mutations in CML.
- Identifying and monitoring these mutations can enhance patient prognosis and survival.
- dHPLC holds potential for routine clinical application in patient testing.
More Related Videos
07:49Characterize Disease-related Mutants of RAF Family Kinases by Using a Set of Practical and Feasible Methods
Published on: July 17, 2019
10:41Wild-type Blocking PCR Combined with Direct Sequencing as a Highly Sensitive Method for Detection of Low-Frequency Somatic Mutations
Published on: March 29, 2017