{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,10]],"date-time":"2026-08-10T13:30:17Z","timestamp":1786368617638,"version":"build-2736575974"},"reference-count":30,"publisher":"MDPI AG","issue":"16","license":[{"start":{"date-parts":[[2026,8,10]],"date-time":"2026-08-10T00:00:00Z","timestamp":1786320000000},"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>Background\/Objectives: Prostate cancer (PCa) and benign prostatic hyperplasia (BPH) are highly prevalent among African men. While single-nucleotide polymorphisms (SNPs) in KLK3, HNF1B, and JAZF1 are established risk markers in Western cohorts, their specific allele frequencies and diagnostic utility remain poorly characterized in African populations. This study evaluated the diagnostic accuracy, sensitivity, and cost-effectiveness of High-Resolution Melting (HRM) PCR for germline SNP genotyping in a cohort of Nigerian men. Methods: Peripheral blood buffy coat DNA was extracted from 31 male participants attending the urology clinic at Lagos University Teaching Hospital, classified into PCa (n = 13), BPH (n = 12), and healthy controls (n = 6). Real-time PCR and HRM analysis were optimized to genotype three specific risk variants: rs2735839 (KLK3), rs4430796 (HNF1B), and rs10486567 (JAZF1). Allelic discrimination was validated using sequence-verified, synthetic positive controls for homozygous wild-type and mutant variants. HRM analysis generated reproducible amplification and melting profiles that enabled clear discrimination of wild-type, heterozygous, and variant genotypes for all three loci. Results: The risk-associated alleles of KLK3 (G), HNF1B (C), and JAZF1 (T) occurred more frequently among patients with prostate cancer than among those with benign prostatic hyperplasia. Clinically, prostate cancer patients also presented with markedly higher PSA concentrations, older age at diagnosis, and higher Gleason scores, consistent with more aggressive disease. The optimized HRM workflow required less than two hours from amplification to genotype assignment and eliminated the need for post-PCR processing, making it suitable for routine molecular testing in resource-constrained laboratories. Conclusions: HRM-based genotyping of KLK3 (rs2735839), HNF1B (rs4430796), and JAZF1 (rs10486567) provides a highly reproducible, rapid, and low-cost molecular tool for mapping prostate disease risk variants. Because it requires no post-PCR processing, this closed-tube HRM workflow is ideally suited for resource-limited clinical settings to enhance early risk stratification and differentiate malignant PCa from benign BPH.<\/jats:p>","DOI":"10.3390\/diagnostics16162521","type":"journal-article","created":{"date-parts":[[2026,8,10]],"date-time":"2026-08-10T12:47:36Z","timestamp":1786366056000},"page":"2521","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["High-Resolution Melting PCR-Based SNP Germline Genotyping of KLK3, JAZF1, and HNF1B in Prostate Cancer and Benign Prostatic Hyperplasia in Nigerian Men\u2014A Pilot Study"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5530-6205","authenticated-orcid":false,"given":"Lateef Adegboyega","family":"Sulaimon","sequence":"first","affiliation":[{"name":"Department of Biochemistry, Faculty of Life Science, University of Abuja, Abuja 900109, Nigeria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ayorinde Babatunde","family":"James","sequence":"additional","affiliation":[{"name":"The GeneLab Bioscience, Yaba, Lagos 101212, Nigeria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aishat Abisola","family":"Akinbami","sequence":"additional","affiliation":[{"name":"Department of Chemical Sciences, College of Natural and Applied Science, Crescent University, Abeokuta 111105, Nigeria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Oladayo Musa","family":"Babalola","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Faculty of Life Science, University of Lagos, Lagos 101017, Nigeria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Grace Folashayo","family":"Oni","sequence":"additional","affiliation":[{"name":"The GeneLab Bioscience, Yaba, Lagos 101212, Nigeria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ganiu Adigun","family":"Idris","sequence":"additional","affiliation":[{"name":"The GeneLab Bioscience, Yaba, Lagos 101212, Nigeria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mayowa","family":"Agbi","sequence":"additional","affiliation":[{"name":"The GeneLab Bioscience, Yaba, Lagos 101212, Nigeria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Akintayo Ashraf","family":"Akintola","sequence":"additional","affiliation":[{"name":"Department of Biology, Teachers College and Institute for Phylogenomics and Evolution, Kyungpook National University, Daegu 41566, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Modinat Olawunmi","family":"Lawal","sequence":"additional","affiliation":[{"name":"Department of Chemical Sciences, College of Natural and Applied Science, Crescent University, Abeokuta 111105, Nigeria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adewale Segun","family":"James","sequence":"additional","affiliation":[{"name":"School of Integrated Biomedical Sciences, USF Morsani College of Medicine, University of South Florida, Tampa, FL 33620, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,8,10]]},"reference":[{"key":"ref_1","first-page":"7","article-title":"Cancer statistics, 2021","volume":"71","author":"Siegel","year":"2021","journal-title":"CA Cancer J. Clin."},{"key":"ref_2","first-page":"394","article-title":"Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries","volume":"68","author":"Bray","year":"2018","journal-title":"CA Cancer J. Clin."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1489","DOI":"10.1016\/S1470-2045(19)30625-4","article-title":"Navigating prostate cancer control in Nigeria","volume":"20","author":"Chidebe","year":"2019","journal-title":"Lancet Oncol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1148","DOI":"10.1016\/j.sjbs.2020.02.003","article-title":"A piece in prostate cancer puzzle: Future perspective of novel molecular signatures","volume":"27","author":"Nassir","year":"2020","journal-title":"Saudi J. Biol. Sci."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"437","DOI":"10.1038\/gim.2012.182","article-title":"How can polygenic inheritance be used in population screening for common diseases?","volume":"15","author":"Khoury","year":"2013","journal-title":"Genet. Med."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"D896","DOI":"10.1093\/nar\/gkw1133","article-title":"The new NHGRI-EBI Catalog of published genome-wide association studies (GWAS Catalog)","volume":"45","author":"MacArthur","year":"2017","journal-title":"Nucleic Acids Res."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"166","DOI":"10.1002\/pros.24635","article-title":"Evaluation of HNF1B, KLK3, ELAC2, TMPRSS2-ERG, and CTNNB1 polymorphisms associated with prostate cancer in samples of patients from HUPE-UERJ","volume":"84","author":"Menezes","year":"2024","journal-title":"Prostate"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"601","DOI":"10.1002\/pros.21094","article-title":"HNF1B and JAZF1 genes, diabetes, and prostate cancer risk","volume":"70","author":"Stevens","year":"2010","journal-title":"Prostate"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"962","DOI":"10.1038\/labinvest.2015.73","article-title":"Transcriptional upregulation of HNF-1\u03b2 by NF-\u03baB in ovarian clear cell carcinoma modulates susceptibility to apoptosis through alteration in Bcl-2 expression","volume":"95","author":"Suzuki","year":"2015","journal-title":"Lab. Investig."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1016\/j.semradonc.2016.08.002","article-title":"Prostate cancer genetics: Variation by race, ethnicity, and geography","volume":"27","author":"Rebbeck","year":"2017","journal-title":"Semin. Radiat. Oncol."},{"key":"ref_11","first-page":"869","article-title":"Gene-environment interactions between JAZF1 and occupational and household lead exposure in prostate cancer among African American men","volume":"25","author":"Levin","year":"2014","journal-title":"Cancer Causes Control"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Takano, E.A., Mitchell, G., Fox, S.B., and Dobrovic, A. (2008). Rapid detection of carriers with BRCA1 and BRCA2 mutations using high-resolution melting analysis. BMC Cancer, 8.","DOI":"10.1186\/1471-2407-8-59"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"van Pelt-Verkuil, E., van Belkum, A., and Hays, J.P.H. (2008). Ensuring PCR quality\u2014Laboratory organisation, PCR optimization and controls. Principles and Technical Aspects of PCR Amplification, Springer.","DOI":"10.1007\/978-1-4020-6241-4"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"100014","DOI":"10.1016\/j.arres.2021.100014","article-title":"Mitoquinol mesylate alleviates oxidative damage in cirrhotic and advanced hepatocellular carcinogenic rats through mitochondrial protection and antioxidative effects","volume":"3","author":"Sulaimon","year":"2021","journal-title":"Adv. Redox Res."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Farrar, J.S., and Wittwer, C.T. (2017). High-resolution melting curve analysis for molecular diagnostics. Molecular Diagnostics, Academic Press. [3rd ed.]. Chapter 6.","DOI":"10.1016\/B978-0-12-802971-8.00006-7"},{"key":"ref_16","first-page":"329","article-title":"High-resolution melting (HRM) genotyping","volume":"Volume 2638","author":"Shavrukov","year":"2023","journal-title":"Plant Genotyping"},{"key":"ref_17","unstructured":"SatrioBio (2024). hrmPy (v0.1): High-Resolution Melting Analysis Package, GitHub, Inc.. Available online: https:\/\/github.com\/satriobio\/hrmpy."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Cui, H., Zhang, W., Zhang, L., Qu, Y., Xu, Z., Tan, Z., Yan, P., Tang, M., Yang, C., and Wang, Y. (2024). Risk factors for prostate cancer: An umbrella review of prospective observational studies and Mendelian randomization analyses. PLoS Med., 21.","DOI":"10.1371\/journal.pmed.1004362"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Christelle, B.M.D., Gebreselassie, K.H., Abdelkerim, N.S., Dieudonn\u00e9, Z.O.J., Makon, A.S.N., Kifle, A.T., Mohammed, T.O., Cassell, A.K., Youssef, B., and Idrissa, S. (2025). Prostate cancer diagnosis and management: Current practices in Africa\u2014A consultant-based survey. Front. Urol., 5.","DOI":"10.3389\/fruro.2025.1496951"},{"key":"ref_20","first-page":"159","article-title":"Increased incidence of prostate cancer in Nigerians","volume":"91","author":"Ogunbiyi","year":"1999","journal-title":"J. Natl. Med. Assoc."},{"key":"ref_21","first-page":"231","article-title":"Epidemiology of prostate cancer in Nigeria: A mixed methods systematic review","volume":"36","author":"Iheanacho","year":"2025","journal-title":"Cancer Causes Control"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"66","DOI":"10.1186\/s12301-024-00470-x","article-title":"Prevalence of benign prostatic hyperplasia and prostate cancer among men in Cross River State, Nigeria: A sonographic study","volume":"30","author":"Eze","year":"2024","journal-title":"Afr. J. Urol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"153","DOI":"10.4103\/aam.aam_101_20","article-title":"Prostate cancer characteristics: A descriptive analysis of clinical features at presentation in the last decade in a Black African community","volume":"21","author":"Ogunbiyi","year":"2022","journal-title":"Ann. Afr. Med."},{"key":"ref_24","first-page":"238","article-title":"Outcome of radical retropubic prostatectomy at the Lagos State University Teaching Hospital","volume":"57","author":"Adebayo","year":"2023","journal-title":"Niger. Med. J."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Khaled, K., Jardaly, H., and Almilaji, O. (2025). Prostate Cancer and Dietary Sugar Intake: A Systematic Review. Onco, 5.","DOI":"10.3390\/onco5030031"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1186\/s12301-023-00365-3","article-title":"Quantifying the changes in body mass index and performance status during androgen deprivation therapy for advanced prostate cancer in southeast Nigeria","volume":"29","author":"Nnabugwu","year":"2023","journal-title":"Afr. J. Urol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"9587","DOI":"10.1038\/s41467-024-52472-6","article-title":"A PSA SNP associates with cellular function and clinical outcome in men with prostate cancer","volume":"15","author":"Srinivasan","year":"2024","journal-title":"Nat. Commun."},{"key":"ref_28","first-page":"2958","article-title":"Epigenetic silencing of HNF1B in prostate cancer correlates with tumor aggressiveness and poor prognosis","volume":"184","author":"Gao","year":"2010","journal-title":"J. Urol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"767","DOI":"10.1080\/15257770.2021.1944636","article-title":"PHLPP2 gene L1016S (rs61733127) and PIK3R1 gene Met326Ile (rs3730089) polymorphisms are associated with the risk of colon and breast cancers","volume":"40","author":"Daneshvar","year":"2021","journal-title":"Nucleosides Nucleotides Nucleic Acids"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Jafari, P., Baghernia, S., Moghanibashi, M., and Mohamadynejad, P. (2022). Significant association of variable number tandem repeat polymorphism rs58335419 in the MIR137 gene with the risk of gastric and colon cancers. Br. J. Biomed. Sci., 79.","DOI":"10.3389\/bjbs.2021.10095"}],"container-title":["Diagnostics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2075-4418\/16\/16\/2521\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,10]],"date-time":"2026-08-10T12:50:54Z","timestamp":1786366254000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2075-4418\/16\/16\/2521"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,10]]},"references-count":30,"journal-issue":{"issue":"16","published-online":{"date-parts":[[2026,8]]}},"alternative-id":["diagnostics16162521"],"URL":"https:\/\/doi.org\/10.3390\/diagnostics16162521","relation":{},"ISSN":["2075-4418"],"issn-type":[{"value":"2075-4418","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,10]]}}}