Simultaneous detection of fungal (1,3)-β-d-glucan and procalcitonin using a dual-label time-resolved fluorescence immunoassay

摘要

Neonatal infectious diseases are a serious threat to the health of newborns. The aim was to establish a new detection method for the simultaneous measurement of (1,3)-beta-d-glucan and procalcitonin in serum for the early screening and efficacy testing of neonatal infectious diseases. We established a sandwich dual-label time-resolved fluorescence immunoassay (TRFIA): anti-(1,3)-beta-d-glucan/procalcitonin antibodies immobilized on 96-well plates captured (1,3)-beta-d-glucan/procalcitonin antigens and then banded together with the detection antibodies labeled with europium(III) (Eu3+)/samarium(III) (Sm3+) chelates. Finally, time-resolved fluorometry was used to measure the fluorescence intensity. The linear correlation coefficient (R-2) of the (1,3)-beta-d-glucan standard curve was 0.9913, and the R-2 of the procalcitonin standard curve was 0.9911. The detection sensitivity for (1,3)-beta-d-glucan was 0.4 pg/mL (dynamic range: 0.6-90 pg/mL), and the average recovery was 101.55%. The detection sensitivity for procalcitonin was 0.02 ng/mL (dynamic range: 0.05-95 ng/mL), and the average recovery was 104.61%. There was a high R-2 between the present TRFIA method and a commercially available assay (R-2 = 0.9829 for (1,3)-beta-d-glucan and R-2 = 0.9704 for procalcitonin). Additionally, the cutoff values for (1,3)-beta-d-glucan and procalcitonin were 23.95 pg/mL and 0.055 ng/mL, respectively. The present TRFIA method has high sensitivity, accuracy, and specificity and is an effective method for early screening and efficient testing of neonatal invasive fungal infection.

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