Raman Spectroscopy of Cell-Free Cervicovaginal Lavage for HPV Lesion Diagnosis: A Pilot Study

High-risk human papillomavirus (HPV) is the leading etiological factor in cervical cancer, creating a pressing need for less invasive and more objective diagnostic tools. This pilot study pioneers the application of Raman spectroscopy to cell-free cervicovaginal lavage (CVL) for distinguishing between low-grade and high-grade squamous intraepithelial lesions (LSIL and HSIL) in HPV-positive patients. Raman spectra were acquired at 532-nm excitation from cell-free CVL samples of 20 patients with histologically confirmed LSIL (<i>n</i> = 9) or HSIL (<i>n</i> = 11). Comparative analysis of Raman bands revealed a significant biochemical shift in HSIL, presumably characterized by reduced glycogen and lactate/lactic acid levels alongside substantially elevated heme proteins. A diagnostic model based on key spectral intensity ratios achieved differentiation between LSIL and HSIL with 80% sensitivity and 86% specificity. These findings demonstrate that Raman spectroscopy of cell-free CVL effectively captures profound metabolic and microvascular alterations characteristic of neoplastic progression, showcasing its strong potential as a rapid, cost-effective, non-invasive, and objective tool for cervical lesion risk stratification.