Comparative performance of standalone E6/E7 mRNA testing versus a combined DNA genotyping and viral load assay for...

To compare the diagnostic performance of standalone E6/E7 mRNA testing versus a combined HPV DNA genotyping and viral load assay for detecting cervical high-grade lesions (CIN2+) and to validate performance, biological rationale, and clinical utility.

This single-center, retrospective, paired diagnostic accuracy study included 150 women in the primary cohort and 100 in an independent validation cohort, all referred for colposcopy. Participants underwent both the E6/E7 mRNA test (Aptima) and the combined DNA test (genotyping + Hybrid Capture 2 viral load) within one month prior to biopsy. Histopathology was the reference standard. Analyses included diagnostic performance metrics, correlation of viral load with histopathology, subgroup assessments, and follow-up of 65 DNA+/mRNA- women.

In the primary cohort, the sensitivity of the mRNA assay (87.7%) was not significantly different from that of the DNA test (94.7%, P=0.221), but the specificity was significantly higher (P<0.001). The mRNA signal strongly correlated with histopathological severity (ρ=0.72, P<0.001) and was stronger than the DNA load (ρ=0.38, P<0.001). Validation confirmed these results. Subgroup analyses showed superior mRNA specificity in women <30 years (96.7% vs. 60.0%) and for ASCUS triage (92.7% vs. 63.4%). The mRNA assay effectively refined risk among DNA-positive patients.

The E6/E7 mRNA assay provides significantly higher specificity and positive predictive value for CIN2+ detection than a comprehensive DNA-based strategy, without significant sensitivity loss. Its reproducible performance, biological rationale, and proven safety support its role in enhancing precision screening, particularly for ASCUS, young women, and HPV DNA-positive individuals.