Introduction: Successful medical decisions are highly reliant on medical laboratory results; as such, laboratory results are essential to patient health management, and errors in those results may lead to serious complications and death. The aim of the present study was to assess the knowledge of medical laboratory practitioners in Saudi Arabia regarding factors that may influence medical laboratory testing results and the correct actions for troubleshooting. Methods: A total of 256 practitioners participated, representing diverse qualifications and various healthcare sectors. The study utilized a pretested questionnaire comprised of three sections: socio-demographic characteristics, knowledge, and attitudes. Results: The responses demonstrated participants to possess outstanding knowledge of factors that may affect laboratory test results, including biological and pathological factors, diet, and personal habits. Furthermore, participants demonstrated satisfactory attitudes toward such factors. Conclusions: Thus, this study reveals a magnificent image concerning Saudi practitioners’ knowledge and capacity for suitable actions given unexpected test results, and hence their ability to provide correct healthcare services. This constitutes an acceptable overview, particularly as a first pass; nonetheless, more studies on larger cohorts remain required.