BackgroundTotal laboratory automation (TLA) has the potential to reduce specimen processing time, optimize workflow, and decrease turnaround time (TAT). The purpose of this research is to investigate whether the TAT of our laboratory has changed since the adoption of TLA, as well as to optimize laboratory workflow, improve laboratory testing efficiency, and provide better services of clinical diagnosis and treatment.Materials and MethodsLaboratory data was extracted from our laboratory information system in two 6-month periods: pre-TLA (July to December 2019) and post-TLA (July to December 2020), respectively.ResultsThe median TAT for positive cultures decreased significantly from pre-TLA to post-TLA (65.93 vs 63.53, P<0.001). For different types of cultures, The TAT of CSF changed the most (86.76 vs 64.30, P=0.007), followed by sputum (64.38 vs 61.41, P<0.001), urine (52.10 vs 49,57, P<0.001), blood (68.49 vs 66.60, P<0.001). For Ascites and Pleural fluid, there was no significant difference (P>0.05). Further analysis found that the incidence of broth growth only for pre-TLA was 12.4% (14/133), while for post-TLA, it was 3.4% (4/119). The difference was statistically significant (P=0.01). The common isolates from CSF samples were Cryptococcus neoformans, coagulase-negative Staphylococcus, Acinetobacter baumannii, and Klebsiella pneumonia.ConclusionUsing TLA and setting up three shifts shortened the TAT of our clinical microbiology laboratory, especially for CSF samples.