Minimal Residual Disease Detection Using Gene Scanning Analysis, Fluorescent Fragment Analysis, and Capillary Electrophoresis for IgH Rearrangement in Adult B-Lineage Acute Lymphoblastic Leukemia: A Cross-Sectional Study

Abstract

Objective: Minimal residual disease (MRD) is considered the greatest prognostic factor in acute lymphoblastic leukemia(ALL). MRD is a valuable tool for anticipating impending relapse and treatment response assessment. The objective ofthe present study was to investigate whether the detection of IgH gene rearrangement using polymerase chain reaction(PCR)-based GeneScan analysis could be a complementary method to monitor MRD along with the quantitative realtimePCR (qPCR).Materials and Methods: In this cross-sectional study, we valued the MRD levels, based on the GeneScanning analysis(GSA), and then compared the data with quantitative real-time polymerase chain reaction at different time points inperipheral blood (PB) samples of adult B-lineage ALL patients (n=35). The specific polymerase chain reaction (PCR)primers for IGH gene FR-1 and fluorescence-labeled J-primer were used and analyzed by capillary gel electrophoresison a sequencer. The results of this study were compared with the previously reported MRD results obtained by the IGHrearrangements allele-specific oligonucleotide (ASO) -qPCR methods.Results: The total concordance rate was 86.7%, with a P<0.001. MRD results obtained by GSA and ASO-qPCR methodswere concordant in all diagnostic samples and samples on the 14th and 28th days of induction therapy. The results of these 2.5years’ follow-ups demonstrated a significant correlation between the two techniques (r=0.892, P<0.001).Conclusion: It seems that the PCR-based GeneScan analysis of IGH gene rearrangement detection may be a valuablemolecular technique to distinguish monoclonality from polyclonality. And also, it may be a precise tool to detect theresidual leukemic DNA in the PB follow-up samples of patients.

Keywords

• acute lymphoblastic leukemia
• capillary gel electrophoresis
• immunoglobulin heavy chain
• genescanning
• minimal residual disease