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Background: Developmental dyslexia is one of the most common neurobehavioral disorders affecting children, but prevalence data on this condition are poor. The objective of the present study is to determine the prevalence of dyslexia in Italy in an unselected school population, using clearly defined diagnostic criteria and methods. Methods: Cross-sectional study carried out in nine Italian Regions: two located in Northern Italy (Friuli Venezia Giulia and Veneto), three in Central Italy (Marche, Lazio and Umbria) and four in Southern Italy (Abruzzo, Molise, Puglia and Sardegna). Three consecutive levels of screening were carried out: the first two at school, to screen the population and identify children with suspect dyslexia; the last in centers with multi-professional staff specialized in learning disabilities to confirm the diagnosis. The key outcome measure is the prevalence of dyslexia, defined as the ratio between the number of children confirmed positive at the third level of screening and the total number of children enrolled in the study. Results: We finally recruited 11094 children aged 8–10 years, of which 9964 constituted the final working sample after applying exclusion criteria and including only children who received parents’ consent to participate. The prevalence of dyslexia in the whole sample was 3.5% (95% CI 3.2–3.9%), with little differences between Northern, Central and Southern Italy (respectively 3.6%, 3.2% and 3.7%). In almost two out of three children with dyslexia the disorder had not been previously diagnosed. Conclusions: This study confirms that in primary school children at the age of 8–10 years in Italy dyslexia is widely underestimated. Reliable data on dyslexia prevalence are needed to allocate necessary human and financial resources both to Health Services and Schools, ensuring timely support to children and families. [ABSTRACT FROM AUTHOR]