Bloodstain pattern analysis (BPA) has long been accepted by courts as an area of expertise; however, that position has recently been challenged. The discipline has been criticized for limited empirical research into practitioner determination error rates and whether determinations require specialized knowledge/expertise, including whether practitioner experience level influences accuracy. This study attempted to address these knowledge gaps as they relate to bloodstain pattern recognition. The aims were twofold: to establish whether practitioners would outperform lay non-practitioners, and whether practitioner experience influenced accuracy and error in determinations. Comparisons of practitioner responses under three scenarios (forced, casework, and definitive) were also made to assess conservatism/certainty in pattern recognition. Participants (both BPA practitioners and non-practitioners) analyzed photographs of bloodstain patterns and made determinations of the broad bloodstain category and specific bloodstain pattern type. When forced to provide only a single response, practitioners identified bloodstain categories and patterns significantly more accurately than non-practitioners (p = 0.0001, p < 0.00001, respectively). Practitioner accuracy in bloodstain pattern recognition was positively associated with experience level (p = 0.0429) and this was consistent regardless of response scenario. Although no significant difference in practitioner accuracy was observed across response scenarios, practitioner conservatism/certainty varied significantly among the broad bloodstain category and specific pattern types. Overall, these results support bloodstain pattern recognition as an area of expertise and that practitioner experience positively influences accuracy. Based on these results, a series of recommendations were proposed aimed at further improving practices within the discipline to maximize accuracy and reliability of BPA evidence.