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Simultaneous detection of fungal (1,3)‐β‐ d ‐glucan and procalcitonin using a dual‐label time‐resolved fluorescence immunoassay
Author(s) -
Li Huitao,
Huang Zhifeng,
Lin Binchun,
Chen Xueyu,
Xiong Xiaoyun,
Cao Aifen,
Yang Chuanzhong
Publication year - 2021
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1002/bab.1908
Subject(s) - procalcitonin , immunoassay , detection limit , chemistry , standard curve , chromatography , glucan , beta glucan , antibody , medicine , microbiology and biotechnology , immunology , biology , biochemistry , sepsis
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)‐β‐ 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)‐β‐ d ‐glucan/procalcitonin antibodies immobilized on 96‐well plates captured (1,3)‐β‐ d ‐glucan/procalcitonin antigens and then banded together with the detection antibodies labeled with europium(III) (Eu 3+ )/samarium(III) (Sm 3+ ) chelates. Finally, time‐resolved fluorometry was used to measure the fluorescence intensity. The linear correlation coefficient ( R 2 ) of the (1,3)‐β‐ 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)‐β‐ 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)‐β‐ d ‐glucan and R 2 = 0.9704 for procalcitonin). Additionally, the cutoff values for (1,3)‐β‐ 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.