Abstract Background The pre-participation screening is crucial as the risk of sudden death for competitive athletes is higher than that of non-athletes. In Turkey, screening could be performed at the primary health care setting both by family medicine specialists (FMSs) and by general practitioners (GPs). Although there is a guideline, there is no legal regulation for the process. The aim of this study is to evaluate the approach of primary care physicians in pre-participation screening. Methods An online questionnaire based on the Turkish Medical Association (TMA) guideline, and the 14-item Screening Guideline of the American Heart Association (AHA) was delivered to primary care physicians. Knowledge, experience and approach of the physicians were assessed. Self-confidence, application of AHA criteria, accurate referral ratios and further test requests were taken as outcome measures. Descriptive and inferential analyzes were performed and p < 0.05 was considered significant. Results Of 214 participants, 39.3% were women, the average age was 44.9 years, the average work experience was 7.9 years. 89.7% of the participants were aware of their authorization, 90.2% had previously given this report, but only 6.5% feels confident. Only 13.1% knows the presence of TMA guideline, and only 23.8% states being educated on the subject at any part of their career. More than 60% of the participants consider further testing necessary in addition to medical history and physical examination. Blood and urine test requests were significantly more for GPs compared to FMSs (p = 0.026, p = 0.011). Accurate referral decision ratio was only 59.3% with no difference between FMSs and GPs (p = 0.216). Work experience had no effect on any of the outcomes. Conclusions As the legal regulations and awareness of the guideline are insufficient, pre-participation screening is not standardized, and this increases tendency for further testing or referral. Key messages The pre-participation screening of athletes in Turkey is not standardized among primary care physicians. Further testing request ratio is high and accurate referral ratio is low.