ABSTRACT
The presence of the BCR-ABL1 transcript is essential in establishing a diagnosis of Chronic Myeloid Leukaemia (CML). In the absence of BCR-ABL1 , JAK2 V617F mutation testing is performed to support the diagnosis of Polycythemia Vera (PV), Essential Thrombocythemia (ET), and primary myelofibrosis (MF). We wanted to test the feasibility of multiplex detection of BCR-ABL1 transcript variants and JAK2 V617F mutation simultaneously using the Reverse Dot-Blot Hybridisation (RDB) method. The Limit of Detection (LOD) or analytical sensitivity of the RDB method using cDNA specimens was 0.5% and 6.25% in detecting BCR-ABL1 and JAK2 mutant transcripts, respectively. Diagnostic specificity and sensitivity to detect BCR ABL1 and JAK2 were 100% and 85.7%; 92% and 100%, respectively. RDB also detected BCR-ABL1 transcripts in 22% of JAK2 V617F mutation positive samples (N = 36). RT-PCR RDB is a promising qualitative multiplexing method to detect simultaneous BCR-ABL1 and JAK2 mutant transcripts.
