Development of a Highly Sensitive β-Glucan Detection System Using Scanning Single-Molecule Counting Method.

Autor: Adachi, Yoshiyuki, Nakata, Hidetaka, Tanabe, Tetsuya, Yamanaka, Daisuke, Kanno, Takashi, Ishibashi, Ken-ichi, Ohno, Naohito
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Zdroj: International Journal of Molecular Sciences; Jun2021, Vol. 22 Issue 11, p5977, 1p
Abstrakt: To overcome the limitations of the Limulus amebocyte lysate (LAL) assay method for the diagnosis of invasive fungal infection, we applied a reaction system combining recombinant β-glucan binding proteins and a scanning single-molecule counting (SSMC) method. A novel (1→3)-β-D-glucan recognition protein (S-BGRP) and a (1→6)-β-glucanase mutant protein were prepared and tested for the binding of (1→6)-branched (1→3)-β-D-glucan from fungi. S-BGRP and (1→6)-β-glucanase mutant proteins reacted with β-glucan from Candida and Aspergillus spp. Although LAL cross-reacted with plant-derived β-glucans, the new detection system using the SSMC method showed low sensitivity to plant (1→3)-β-D-glucan, which significantly improved the appearance of false positives, a recognized problem with the LAL method. Measurement of β-glucan levels by the SSMC method using recombinant β-glucan-binding proteins may be useful for the diagnosis of fungal infections. This study shows that this detection system could be a new alternative diagnostic method to the LAL method. [ABSTRACT FROM AUTHOR]
Databáze: Complementary Index
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