An In-House Method for Molecular Monitoring of BCR-ABL

作者：Sercan Hakki Ogun^*; Ozturk Ilgin; Caliskan Ceyda; Pehlivan Melek; Sercan Zeynep

来源：Turkish Journal of Hematology, 2012, 29(4): 348-353.

DOI：10.5152/tjh.2011.76

• Chronic myeloid leukemia
• BCR-ABL
• Real-time PCR
• Molecular response

Objective: At present, there are a limited number of facilities in Turkey that can provide reliable real-time quantitative (RQ)-PCR BCR-ABL results. The present study aimed to test a cost-effective, in-house method of BCR-ABL quantification, including verification of the method by RQ-PCR validation tests. %26lt;br%26gt;Material and Methods: BCR-ABL and ABL target sequences were cloned into pJET1.2 vectors, from which calibrators were prepared and used as templates in RQ-PCR reactions to generate standard curves. Dilutions of K562 cells (representing an in vitro simulation of BCR-ABL transcript reduction) were analyzed. %26lt;br%26gt;Results: Standard curves were generated from calibrators. These curves were then used to calculate the BCR-ABL and ABL copy numbers; in which linear BCR-ABL results were obtained. Repetitive experiments showed that our methodology was able to detect 1 BCR-ABL positive cell from amnong 1x10(5) cells. %26lt;br%26gt;Conclusion: The method described herein is suitable for implementation with any RQ-PCR instrument and/or kit for quantify BCR-ABL transcripts.

• 出版日期2012-12