{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T20:58:06Z","timestamp":1792789086838,"version":"4.5.0"},"reference-count":36,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2026,9,17]],"date-time":"2026-09-17T00:00:00Z","timestamp":1789603200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Health Directorate of the Lazio Region, Italy","id":[{"id":"https:\/\/ror.org\/008hssd09","id-type":"ROR","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cancers"],"abstract":"<jats:p>ASC-US refers to cytological abnormalities suggestive of squamous intraepithelial lesions (SILs) that are qualitatively or quantitatively insufficient for a definitive diagnosis. In addition to HPV infection, which is the etiological agent of preneoplastic and neoplastic cervical lesions, several other conditions may induce cytological changes classified as ASC-US, including inflammation, atrophy with degeneration, hormonal effects, and technical artifacts. The aim of this study was to evaluate the incidence of primary Pap tests diagnosed as ASC-US that also tested positive for HR-HPV genotypes. In this retrospective study, 6181 primary Pap tests collected between 21 July 2021 and 9 January 2024 from the Roma 2 Local Health District were reviewed. ThinPrep containers (Hologic) were used for liquid-based cytology, allowing both cytological examination and molecular testing for HR-HPV detection. The Anyplex\u2122 II HR HPV Detection assay (Seegene) was used for the identification, by real-time PCR, of 14 HR-HPV genotypes (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68). Among the 6181 Pap tests analyzed, 202 cases (3.3%) were diagnosed as ASC-US and subsequently underwent HR-HPV molecular testing. Of these, 107 cases (53.0%) were negative, while 95 (47.0%) were positive for HR-HPV. In HPV-positive samples, single-genotype infections were detected in 65 cases (68.4%), while multiple genotypes were identified in 30 cases (31.6%). A total of 135 HR-HPV genotypes were detected. HPV 16 was identified in nine cases (6.7%), HPV 18 in six (4.4%), HPV 31 in nine (6.7%), HPV 33 in five (3.7%), HPV 35 in two (1.5%), HPV 39 in 10 (7.4%), HPV 45 in four (3.0%), HPV 51 in 20 (14.8%), HPV 52 in seven (5.2%), HPV 56 in 15 (11.1%), HPV 58 in eight (5.9%), HPV 59 in 17 (12.6%), HPV 66 in 12 (8.9%), and HPV 68 in 11 cases (8.1%). Our findings indicate that approximately half of ASC-US diagnoses are associated with HR-HPV infection. Interestingly, the detected viral genotypes mainly belonged to less common HR-HPV strains. HR-HPV testing following an ASC-US cytological diagnosis is essential for colposcopy triage, while extended genotyping enables the monitoring of genotype persistence over time, a key factor in the development of cervical dysplastic and neoplastic lesions.<\/jats:p>","DOI":"10.3390\/cancers18183022","type":"journal-article","created":{"date-parts":[[2026,9,17]],"date-time":"2026-09-17T11:41:35Z","timestamp":1789645295000},"page":"3022","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Primary Pap Smear Cytological Diagnosis of ASC-US: HR-HPV Genotyping and Follow-Up of HPV-Positive Patients"],"prefix":"10.3390","volume":"18","author":[{"given":"Tiziana","family":"Pisani","sequence":"first","affiliation":[{"name":"Unit\u00e0 Operativa Complessa di Patologia Clinica, Azienda Ospedaliera San Giovanni-Addolorata, 00184 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria","family":"Cenci","sequence":"additional","affiliation":[{"name":"Unit\u00e0 Operativa Complessa di Patologia Clinica, Azienda Ospedaliera San Giovanni-Addolorata, 00184 Rome, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ettore Domenico","family":"Capoluongo","sequence":"additional","affiliation":[{"name":"Unit\u00e0 Operativa Complessa di Patologia Clinica, Azienda Ospedaliera San Giovanni-Addolorata, 00184 Rome, Italy"},{"name":"Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Universit\u00e0 Federico II, 80138 Napoli, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,9,17]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"524","DOI":"10.1016\/S0140-6736(13)62218-7","article-title":"Efficacy of HPV-based screening for prevention of invasive cervical cancer: Follow-up of four European randomised controlled trials","volume":"383","author":"Ronco","year":"2014","journal-title":"Lancet"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1076","DOI":"10.1038\/s41416-025-03161-8","article-title":"Italian guidelines for cervical cancer screening. 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