To assess the diagnostic accuracy of 1,3-β-D-glucan (BDG) assay for diagnosing invasive fungal infections (IFI), we searched the Medline and Embase databases, and studies reporting the performance of BDG assays for the diagnosis of IFI were identified. Our analysis was mainly focused on the cutoff level. Meta-analysis was performed using conventional meta-analytical pooling and bivariate analysis. Our meta-analysis covered 28 individual studies, in which 896 out of 4214 patients were identified as IFI positive. The pooled sensitivity, specificity, diagnostic odds ratio, and area under the summary receiver operating characteristic (AUC-SROC) curve were 0.78 [95% confidence interval (CI), 0.75-0.81], 0.81 (95% CI, 0.80-0.83), 21.88 (95% CI, 12.62-37.93), and 0.8855, respectively. Subgroup analyses indicated that in cohort studies, the cutoff value of BDG at 80 pg/mL had the best diagnostic accuracy, whereas in case-control studies the cutoff value of 20 pg/mL had the best diagnostic accuracy; moreover, the AUC-SROC in cohort studies was lower than that in case-control studies. The cutoff value of 60 pg/mL has the best diagnostic accuracy with the European Organization for Research and Treatment of Cancer/Mycoses Study Group criteria as a reference standard. The 60 pg/mL cutoff value has the best diagnostic accuracy with the Fungitell assay compared to the BDG detection assay. The cutoff value of 20 pg/mL has the best diagnostic accuracy with the Fungitec G-test assay, and the cutoff value of 11 pg/mL has the best diagnostic accuracy with the Wako assay. Serum BDG detection is highly accurate for diagnosing IFIs. As such, 60 pg/mL of BDG level can be used as the best cutoff value to distinguish patients with IFIs from patients without IFI (mainly due to Candida and Aspergillus).