Digital PCR quantification of BCR-ABL for CML diagnosis and monitoring in a LMICs setting

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Project Abstract We propose to develop a low resource setting (LRS) digital PCR (dPCR) instrument to detect BCR-ABL transcripts from small blood samples to identify chronic myeloid leukemia (CML) patients who are eligible for tyrosine kinase inhibitor (TKI) therapy in low- and middle- income countries (LMICs). Despite the success of the targeted TKI Glivec (Gleevec, imatinib) in treating CML in developed countries, the majority of the world's CML patients reside in LMICs with limited access to diagnostic testing and TKI treatment. The Glivec International Patient Assistance Program (GIPAP) is one of the most comprehensive and far-reaching global cancer access programs designed by the Novartis (the manufacturer of Glivec) in partnership with the Max Foundation (TMF) to facilitate access to and distribution of Glivec (imatinib) directly to eligible patients through their providers. One of the major challenges of the implementation of GIPAP program is the lack of diagnostic capabilities in many GIPAP countries which are essential for selecting patients for the TKI therapy, since only patients who are properly diagnosed with Philadelphia chromosome positive CML (Ph+ CML) are eligible to participate in the GIPAP program. We hypothesize that a LRS dPCR instrument to detect the BCR-ABL transcript in a highly sensitive manner will enable more health care institutes in LMICs capabilities to diagnose Ph+ CML and participate in the GIPAP program, as well as monitor patients who are being treated with TKI. We will develop the LRS dPCR instrument by adapting technologies recently developed in our laboratories including a novel digital nucleic acid amplification platform based on a self-digitization (SD) microfluidic chip, which partitions an aqueous sample into tens of thousands of nanoliter volumes suitable for PCR, instruments for efficient sample loading using centrifugal force and signal detection using optical disc (OD, e.g. CD, DVD, Blu-Ray)-styled readers which are highly compatible to the LMICs setting. We propose to leverage these innovative technologies to create a LRS dPCR instrument matching or exceeding the sensitivity and specificity of the current state of the art RT-PCR BCR-ABL fusion transcript assay, with minimal sample preparation and without the need of run to run standards. Such an instrument will have significant impact on improving CML patients' survival in LMICs.