Rapid detection of the BCR::ABL1 fusion gene in chronic myeloid leukemia by RT-RAA combined with a lateral flow strip

Chronic myeloid leukemia (CML) is a myeloproliferative neoplasm characterized by the translocation t(9;22)(q34;q11). This results in the fusion of the BCR and ABL1 genes to produce the BCR ∷ ABL1 gene. Currently, molecular methods for detecting the p210 BCR ∷ ABL1 fusion gene have limitations in terms of complexity and time required for the assay. This study integrated the reverse transcriptase recombinase-aided amplification (RT-RAA) technique with the Nfo probe combined with a lateral flow dipstick assay (LFA) to detect the p210 BCR ∷ ABL1 gene. This study applied RT-RAA combined with LFA to establish a rapid assay for BCR ∷ ABL1 detection. To evaluate the diagnostic performance of the RT-RAA-LFA assay, whole blood samples were analyzed. The diagnostic performance of this method was evaluated by comparing it with RT-PCR and dPCR. The entire RT-RAA-LFA assay required 30 min for nucleic acid amplification at 37 °C and a 10-min visual LFA readout at room temperature. In comparison with the standard RT-PCR, the RT-RAA-LFA assay achieved 100% sensitivity and 100% specificity across 82 clinical blood samples, comprising 19 newly diagnosed CML patients, 38 TKI-treated CML patients and 25 normal subjects. However, when compared with digital PCR, the assay demonstrated 59.09% sensitivity and 100% specificity. The RT-RAA-LFA assay is suitable for the rapid, sensitive, and specific detection of the BCR ∷ ABL1 gene in newly diagnosed CML patients.