Comparison of RT-qPCR and RT-Digital PCR for Detection and Quantification of BCR-ABL1 Transcripts in Chronic Myeloid Leukemia

Abstract

Aim: Chronic myeloid leukemia (CML) is a hematological malignancy in the group of myeloproliferative neoplasms.Philadelphia chromosome, t(9;22)(q34;q11), results in the BCR/ABL1 fusion gene. The Philadelphia chromosome could be detected in almost all CML cases.RT-qPCR method is still the most commonly used method for monitoring BCR/ABL1 fusion.RT-digital PCR method is an alternative in quantitative measurement of BCR-ABL1 fusion, but there is not enough information in the literature yet. It was planned to evaluate and compare of RT-qPCR and RT-digital PCR for detection and quantification of BCR-ABL1 transcripts in CML.

Materials and Methods:Totaly, 39CML patients were performed.Total RNA was extracted with RNA extraction kit (QIAamp RNA Blood Mini Kit). Qiagene Rotor-Gene-Q system was used for RT-qPCR method and QX200™ Droplet Digital™ PCR (ddPCR™) system was used for RT-digital PCR testing.

Results:There was significant difference between the groups in the BCR-ABL1/ABL comparison of the samples (p=0.017) (Table 1).

Conclusion:Although the significant difference between RT-digital PCR and RT-qPCR in detection and quantification of BCR-ABL1 transcripts in CML, RT-digital PCR is not more sensitive in all samples. Therefore, further research is needed to obtain a clear understanding of the effectiveness of RT-digital PCR.