Background Sterilization failure is one of the main reasons for surgical site infections (SSIs). The biological indicator (BI) test is the most reliable test to check sterilization efficiencies. But 48 hours BI test result makes the monitoring process time-consuming. Rapid BI testing can be time demanding in this regard. Therefore, the objective is to determine the importance of rapid BI monitoring for the quality assurance of sterile surgical instruments. Methods This study was conducted in the Labaid Cardiac Hospital, Bangladesh from April 1, 2021, to July 8, 2021. A total of 100 steam and 100 ethylene oxide (EO) rapid BIs and an auto reader incubator were used to conduct this research. Quick BI of steam and EO were used once per day and tested by the auto reader. Later, all the tested BIs were incubated for 48 hours by a conventional incubator to confirm the auto reader's rapid BI test results. Result All the EO BI results were found negative, but the BI was found positive twice in steam sterilization. Surgical items of those two loads were re-sterilized. Again, after checking the BI result, the items were released. All BIs except positive steam rapid BIs were found with no growth after 48 hours of incubation for cross-checking of auto reader results. In positive rapid BI of steam, growth was found after 48 hours of incubation. Conclusion When sterilization failure occurred, process recall could not be possible at that time if rapid BI tests were not performed. So, integration of a rapid BI test with an auto reader can save the patient from critical SSI.