Cervical cancer screening is split between molecular approaches and combined cytology and molecular analysis. Here, we present a non-slide based technology to assess cytologic parameters and molecular biomarkers of cervical cancer in a single, high-throughput assay.
Using the IncellDx HPV OncoTect 3Dx assay performed on a FlowSight Image Cytometer (Merck Millipore Sigma), we combined the standard cervical cytology parameters of nuclear-to-cytoplasmic ratio (N/C ratio) and nuclear area with E6, E7 mRNA and the cell cycle parameters of post G0-G1% and aneuploidy to determine the stage of cytologic abnormality.
The combination of directly measured nuclear-to-cytoplasmic ratio (N/C ratio) and E6, E7 mRNA defined with 97% accuracy the cytology result on 200 liquid-based cytology samples (NILM, LSIL, and HSIL). Of the six discrepant samples, four HSIL samples were downgraded to LSIL and two LSIL samples were more likely to be high grade based on marker expression and proliferation rate as determined by post G0-G1%.
Despite regulatory approval of HPV DNA as a primary screening method, the move away from cytology in the screening algorithm has been slow. A method that combines both molecular methods and cytologic parameters in a highly automated system that does not require slides would be desirable. Here, we present an approach that combines direct measurement of cytologic parameters, molecular analysis, and cell cycle with extremely high accuracy for staging cervical disease.