Folate is a micronutrient essential in a variety of biological processes, and an adequate dietary folate intake seems to play a crucial role in health promotion and disease prevention. The importance of strawberry as a natural food source of folate has been recognised only recently, and few pilot studies have investigated the impact of strawberry intake on human folate status. In this study, firstly, we evaluated the folate content of different commercial varieties (Alba, Irma, Patty, Adria, Sveva) and advanced selections (AN99.78.51; AN94.414.52; AN00.239.55) of strawberry. Significant differences were observed among genotypes, confirming the breeding approach as a reliable tool to increase folate content in strawberry. Secondly, the variety Sveva was selected for a medium-term strawberry consumption study, in order to check if a 2-weeks strawberry intake could have any effects on folate status and plasma homocysteine levels, in healthy subjects. An average 3.4% increase in serum folate was observed, however without any statistical significance, as shown by reference change value of each analyte in each subject. This study should be considered as a first pilot investigation, and further investigations are strongly hoped to evaluate the potential impact of strawberry consumption on human folate status, particularly in the case of a previously diagnosed deficiency.