Introduction: High levels of exam anxiety are evident in healthcare students. Practical exams are an integral part of healthcare profession programs. However, no standardised reliable and valid instrument exists to measure practical exam anxiety in healthcare students. Objective: This study aimed to modify a valid and reliable measure used to examine anxiety in job interviews, for use in practical examinations. We then aimed to examine the psychometric properties of the new modified instrument, now characterized as the Measure of Anxiety in Practical Examinations (MAPE) and determine if any differences in gender, personal history of Generalised Anxiety Disorder (GAD) or family history of GAD impacted MAPE scores. Methods: Exploratory factor analysis using principal component analysis was conducted and Cronbach’s alpha examined internal consistency of the instrument. Results: Most A five factor structure was supported (Performance, Appearance, Behaviour, Communication, and Preparedness) which accounted for 60.6% of the variance in responses. The 25 item modified instrument demonstrated sufficient internal consistency (Cronbach’s alpha = 0.93). Females (p = 0.01) and those with a personal history of GAD (0.002) presented with higher MAPE scores. Conclusion: This The MAPE is an acceptable measure of identifying students who present with practical exam anxiety and can help support healthcare profession students to alleviate practical exam anxiety and ensure students’ grades more accurately reflect their skill acquisition. Gender and personal history of GAD can also impact practical exam anxiety and should be considered when addressing practical exam anxiety in healthcare profession students.