Paulina Miranda-Falconi ¹ Gonzalo Flores-Peña ² Mauro F. Jiménez-Trejo ² Yazmin E. Torres-Paz ² Diego O. Reyes-Hernández ² Juan C. Estrada-Guzmán ² Ernesto Hernández-Ramírez ² Erick N. Torres-Torralba ² Juan P. Rangel-Ordoñez ² Daniela K. Vejar-Galicia ² Patricia Reyes-Fonseca ² Omar P. Islas-Bayona ² Rodolfo Hernández-Paredes ² Mercedes Gutiérrez ³ Orlando Santillán ^3*

• ¹ Ginequito Hospital, Monterrey, Nuevo Leon, Mexico
• ² TIMSER Group, Mexico City, Mexico
• ³ ATSO Pharma, Mexico City, Mexico

Cervical cancer is a significant public health issue in Mexico and many developing countries. Early detection is crucial for combating this disease. The official screening test for cervical cancer is cytology, but this technique faces several barriers, including methodological, educational, and sociocultural challenges. Liquid-based cytology is an improved version of this test, however it does not address the aforementioned complications. Biomarkers for cervical precursor lesions and cervical cancer can improve timely detection of the disease. A previous study from our group identified four circulating human proteins as potential biomarkers for these conditions. For molecular screening, we selected GAPDH as the biomarker for cervical precursor lesions and HNRNPA1 as the biomarker for cervical cancer -chosen from the three previously identified options based on antibody availability-to be detected in sera. Participants underwent a comprehensive panel of tests, including liquid-based cytology, PCR detection of Human papillomavirus (HPV), colposcopy, and histopathology -when applicable-. The last two tests were used as references for determining sensitivity and specificity, with histopathology being the gold standard for cervical cancer diagnosis. All the participants successfully received colposcopies (n = 99) and only those women with visible or suspected cervical lesions/malignancies were biopsied (n = 62). A subset of randomly selected biopsies underwent p16 INK4a immunohistochemistry (n = 36). This study compares the performance of liquid-based cytology with the molecular screening. With colposcopy as reference, liquid-based cytology showed 30% sensitivity and 96% specificity, while the molecular screening showed 90% sensitivity and 43% specificity. With histopathology as reference, liquid-based cytology showed 21% sensitivity and 93% specificity, while the molecular screening showed 85% sensitivity and 61% specificity. The molecular screening outperformed the liquid-based cytology in several areas, including detecting true-positive cases, reducing false-negative cases by 34.62%, application time, simplicity of result´s categories, and acceptance among participants. An ideal screening test requires high sensitivity, maintains moderate specificity, and minimizes false negatives. Our proposed screening test meets these criteria, making it an ideal complement -or alternative-for cervical cancer screening.