{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,13]],"date-time":"2026-07-13T08:09:41Z","timestamp":1783930181995,"version":"3.55.0"},"reference-count":28,"publisher":"MDPI AG","issue":"14","license":[{"start":{"date-parts":[[2026,7,11]],"date-time":"2026-07-11T00:00:00Z","timestamp":1783728000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Diagnostics"],"abstract":"<jats:p>Purpose: Severe spermatogenic impairment, including azoospermia, oligozoospermia, asthenozoospermia, and teratozoospermia, represents a major reproductive health concern, yet no validated screening tool exists for early risk stratification in prepubertal boys. We aimed to develop a five-item evidence-based questionnaire and conduct preliminary clinical evaluation to identify prepubertal boys at risk of developing severe spermatogenic impairment. Methods: This retrospective observational study analyzed medical records of 200 male patients aged 18 years who underwent their first semen analysis between 2014 and 2024 at the University of Catania, Italy. Prepubertal risk factor profiles were retrospectively reconstructed by two independent andrologists blinded to semen analysis results across five evidence-based domains: genetic anomalies (0\u20135 points), cryptorchidism (0\u20136 points), gonadotoxic cancer therapy (0\u20137 points), varicocele (0\u20134 points), and hormonal\/endocrine disorders (0\u20135 points), yielding a total score of 0\u201327 points. Scoring weights were based on literature evidence and expert consensus. Reliability was assessed using Cronbach\u2019s \u03b1 (n = 200), test\u2013retest intraclass correlation coefficient (ICC, n = 30, 2-week interval), and inter-rater Cohen\u2019s \u03ba (n = 50). Receiver operating characteristic (ROC) curve analysis identified optimal cut-offs. Semen analysis outcomes were classified according to WHO 2021 criteria. Results: The newly developed questionnaire demonstrated acceptable internal consistency (Cronbach\u2019s \u03b1 = 0.81), good test\u2013retest reliability (ICC = 0.89, 95% CI 0.84\u20130.93), and excellent inter-rater reliability (Cohen\u2019s \u03ba = 0.87). Mean questionnaire scores showed a direct progressive correlation with severity of spermatogenic impairment. The highest scores were observed in azoospermia (mean 14), complete asthenozoospermia (13), and complete teratozoospermia (13). ROC curve analysis identified four clinically meaningful cut-offs: \u22656 (AUC = 0.85, 95% CI 0.79\u20130.91, Youden index = 0.81), \u22657 (AUC = 0.88, 95% CI 0.83\u20130.93, Youden = 0.80), \u226511 (AUC = 0.91, 95% CI 0.87\u20130.95, Youden = 0.79), and \u226513 (AUC = 0.94, 95% CI 0.90\u20130.97, Youden = 0.87). Four risk strata were defined: LOW (0\u20136), MEDIUM (7\u201310), HIGH (11\u201312), and VERY HIGH (\u226513 points). Conclusions: The five-item prepubertal risk questionnaire undergoing preliminary clinical evaluation demonstrates strong correlation with subsequent spermatogenic outcomes, enabling early identification of at-risk boys. The four-tier stratification system provides actionable guidance for clinical management and fertility preservation counseling. However, this cohort was enriched for pathological outcomes due to exclusion of subjects with no known risk factors; cut-offs may not apply to general population screening. Multicenter validation studies in unselected populations are warranted.<\/jats:p>","DOI":"10.3390\/diagnostics16142171","type":"journal-article","created":{"date-parts":[[2026,7,13]],"date-time":"2026-07-13T07:28:27Z","timestamp":1783927707000},"page":"2171","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Development and Preliminary Clinical Evaluation of a Five-Item Prepubertal Risk Questionnaire for Severe Spermatogenic Impairment: A Retrospective Validation Study\u2014Prepubertal Risk Questionnaire for Spermatogenic Impairment"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7113-2372","authenticated-orcid":false,"given":"Sandro","family":"La Vignera","sequence":"first","affiliation":[{"name":"Department of Clinical and Experimental Medicine, University of Catania, Via S. Sofia 78, 95123 Catania, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5217-9343","authenticated-orcid":false,"given":"Rosita A.","family":"Condorelli","sequence":"additional","affiliation":[{"name":"Department of Clinical and Experimental Medicine, University of Catania, Via S. Sofia 78, 95123 Catania, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,7,11]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1186\/s12958-015-0032-1","article-title":"A unique view on male infertility around the globe","volume":"13","author":"Agarwal","year":"2015","journal-title":"Reprod. Biol. Endocrinol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"10-4103","DOI":"10.4103\/aja202552","article-title":"Current risk factors for male infertility and semen parameters: An umbrella review of systematic reviews and meta-analyses","volume":"28","author":"Wang","year":"2026","journal-title":"Asian J. Androl."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"e28218","DOI":"10.4161\/spmg.28218","article-title":"Review of azoospermia","volume":"4","author":"Wosnitzer","year":"2014","journal-title":"Spermatogenesis"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"369","DOI":"10.1038\/s41585-018-0003-3","article-title":"Genetics of male infertility","volume":"15","author":"Krausz","year":"2018","journal-title":"Nat. Rev. Urol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1180","DOI":"10.1016\/j.fertnstert.2013.08.010","article-title":"Effects of chemotherapy and radiotherapy on spermatogenesis in humans","volume":"100","author":"Meistrich","year":"2013","journal-title":"Fertil. Steril."},{"key":"ref_6","unstructured":"Nieschlag, E., Behre, H.M., and Nieschlag, S. (2010). Male Reproductive Health and Dysfunction, Springer. [3rd ed.]."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"152","DOI":"10.1210\/jc.2004-1469","article-title":"Genetic abnormalities among severely oligospermic men who are candidates for intracytoplasmic sperm injection","volume":"90","author":"Foresta","year":"2005","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"734","DOI":"10.1016\/S1472-6483(10)60677-3","article-title":"Male infertility: Role of genetic background","volume":"14","author":"Ferlin","year":"2007","journal-title":"Reprod. Biomed. Online"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"324","DOI":"10.1016\/j.eururo.2012.04.048","article-title":"European Association of Urology guidelines on male infertility: The 2012 update","volume":"62","author":"Jungwirth","year":"2012","journal-title":"Eur. Urol."},{"key":"ref_10","first-page":"S11","article-title":"Selective estrogen receptor modulators and infertility","volume":"55","author":"Hampl","year":"2006","journal-title":"Physiol. Res."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1093\/humupd\/dmm027","article-title":"Epidemiology and pathogenesis of cryptorchidism","volume":"14","author":"Virtanen","year":"2008","journal-title":"Hum. Reprod. Update"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"4588","DOI":"10.1210\/jc.2012-2325","article-title":"Boys with undescended testes: Endocrine, volumetric and morphometric studies on testicular function before and after orchidopexy at nine months or three years of age","volume":"97","author":"Kollin","year":"2012","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"383","DOI":"10.1038\/ng0895-383","article-title":"Diverse spermatogenic defects in humans caused by Y chromosome deletions encompassing a novel RNA-binding protein gene","volume":"10","author":"Reijo","year":"1995","journal-title":"Nat. Genet."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1295","DOI":"10.1016\/S0140-6736(14)60495-5","article-title":"Fertility preservation in men with cancer","volume":"384","author":"Tournaye","year":"2014","journal-title":"Lancet"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"716","DOI":"10.1093\/humrep\/dead004","article-title":"Childhood cancer and hematological disorders negatively affect spermatogonial quantity at diagnosis: A retrospective study of a male fertility preservation cohort","volume":"38","author":"Masliukaite","year":"2023","journal-title":"Hum. Reprod."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Lambertini, M., Del Mastro, L., Pescio, M.C., Andersen, C.Y., Azim, H.A., Peccatori, F.A., Costa, M., Revelli, A., Salvagno, F., and Gennari, A. (2016). Cancer and fertility preservation: International recommendations from an expert meeting. BMC Med., 14.","DOI":"10.1186\/s12916-015-0545-7"},{"key":"ref_17","first-page":"671","article-title":"Fertility preservation in boys facing gonadotoxic cancer therapy","volume":"18","author":"Jensen","year":"2021","journal-title":"Nat. Rev. Urol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"hoae010","DOI":"10.1093\/hropen\/hoae010","article-title":"A 20-year overview of fertility preservation in boys: New insights gained through a comprehensive international survey","volume":"2024","author":"Duffin","year":"2024","journal-title":"Hum. Reprod. Open"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"613","DOI":"10.1016\/S0015-0282(16)55809-9","article-title":"Loss of fertility in men with varicocele","volume":"59","author":"Gorelick","year":"1993","journal-title":"Fertil. Steril."},{"key":"ref_20","unstructured":"World Health Organization (2021). WHO Laboratory Manual for the Examination and Processing of Human Semen, WHO Press. [6th ed.]."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"273","DOI":"10.1016\/S0140-6736(04)16678-6","article-title":"Klinefelter\u2019s syndrome","volume":"364","author":"Lanfranco","year":"2004","journal-title":"Lancet"},{"key":"ref_22","first-page":"167","article-title":"Fertility after bilateral cryptorchidism. Evaluation by paternity, hormone, and semen data","volume":"58","author":"Lee","year":"2002","journal-title":"Horm. Res."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"312","DOI":"10.1093\/humupd\/dmp054","article-title":"Options for fertility preservation in prepubertal boys","volume":"16","author":"Wyns","year":"2010","journal-title":"Hum. Reprod. Update"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"941","DOI":"10.1200\/JCO.1999.17.3.941","article-title":"Semen quality and reproductive hormones before orchiectomy in men with testicular cancer","volume":"17","author":"Petersen","year":"1999","journal-title":"J. Clin. Oncol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"691","DOI":"10.1590\/S1807-59322011000400026","article-title":"An update on the clinical assessment of the infertile male","volume":"66","author":"Esteves","year":"2011","journal-title":"Clinics"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1157","DOI":"10.1111\/j.1464-410X.2010.09959.x","article-title":"Review of current varicocelectomy techniques and their outcomes","volume":"108","author":"Diegidio","year":"2011","journal-title":"BJU Int."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1574","DOI":"10.1007\/s43032-020-00357-6","article-title":"Histologic analysis of testes from prepubertal patients treated with chemotherapy associates impaired germ cell counts with cumulative doses","volume":"28","author":"Medrano","year":"2021","journal-title":"Reprod. Sci."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Puia, C., Iv\u0103nu\u021b\u0103, D., Corlade-Andrei, M., B\u00ee\u021b\u0103, B., and Doroftei, B. (2025). Anti-M\u00fcllerian hormone as a biomarker for predicting testicular sperm extraction outcomes in azoospermic patients: A comprehensive systematic review and meta-analysis. Int. J. Mol. Sci., 26.","DOI":"10.3390\/ijms262311643"}],"container-title":["Diagnostics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2075-4418\/16\/14\/2171\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,13]],"date-time":"2026-07-13T07:37:09Z","timestamp":1783928229000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2075-4418\/16\/14\/2171"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,11]]},"references-count":28,"journal-issue":{"issue":"14","published-online":{"date-parts":[[2026,7]]}},"alternative-id":["diagnostics16142171"],"URL":"https:\/\/doi.org\/10.3390\/diagnostics16142171","relation":{},"ISSN":["2075-4418"],"issn-type":[{"value":"2075-4418","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,11]]}}}