Ultra-precise quantification of mRNA targets across a broad dynamic range with nanoreactor beads

Precise quantification of molecular targets in a biological sample across wide dynamic range is key requirement many diagnostic procedures, such as monitoring response to therapy or detection measurable residual disease. State the art digital PCR assays provide for four orders magnitude. However are complex and require sophisticated microfluidic tools. Here we present an assay format that enables ultra-precise RNA single measurement more than six The approach based on hydrogel beads free compartmentalization they used nanoreactors reverse transcription, amplification combined real time gene transcripts. We have applied these nanoreactor establishing BCR-ABL1 fusion has been characterized its precision linear range. A comparison new method against conventional RT-PCR analysis (reference method) with clinical samples from patients chronic myeloid leukemia (CML) revealed excellent concordance Pearsons correlation coefficient 0.983 slope 1.08.