Abstract:
Objective: The purpose of the study was to compare results and evaluate
the agreement between the endpoint fluorescence (EPF) method
and quantitative real-time polymerase chain reaction (QPCR) during
molecular monitoring of patients with chronic myeloid leukemia (CML)
receiving treatment.
Materials and Methods: The study was conducted at Molecular Lab
of Riphah International University, Islamabad, Pakistan, from January
2017 to December 2018. A total of 150 blood specimens from 30
patients with CML were analyzed at regular intervals during therapy.
The detection/quantification of transcript mRNA was done simultaneously
using QPCR and the EPF method.
Results: Out of a total of 150 RNA specimens analyzed, 117 (78%)
specimens were positive, whereas 33 (22%) were negative for the transcript
using both methods at various stages of treatment. Strong linear
negative correlations between the cycle threshold and relative fluorescence
unit values were observed with P <.0001 at 0, 3, 6, 9, and
12 months of treatment. No significant difference (P >.05) between the
means of the BCR-ABL percentage was observed in either method at all stages of treatment. The bias between the 2 methods was calculated
as 0.069 ± 3.50, and 95% limits of agreement were 6.92% to
–6.79%.
Conclusion: We found that EPF is a simple method to detect/quantify
BCR-ABL mRNA expression during treatment with comparable results
to QPCR.
