The Effect of Intravenous Gamma-globulin Reagents on the Measurement Results of (1→3)-β-D-glucan

Moro, Hiroshi; Koshio, Nao; Bamba, Yuuki; Koizumi, Takeshi; Cho, Hiromi; Aoki, Nobumasa; Hayashi, Masachika; Tsubata, Chikako; Sakagami, Akiko; Satô, Mizuho; Sakagami, Takuro; Koya, Toshiyuki; Tanabe, Yoshinari; Kikuchi, Toshiaki

doi:10.11150/kansenshogakuzasshi.91.1

Cited by 3 publications

(4 citation statements)

References 5 publications

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“…Moro et al reported the detection of BG in immunoglobulin preparations [ 19 ]. This report stated that several immunoglobulin products react positively in the LAL method and that these immunoglobulin products may be a source of false-positive substances [ 6 , 20 ].…”

Section: Discussionmentioning

confidence: 99%

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

Adachi

Nakata

Tanabe

et al. 2021

IJMS

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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.

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Section: Discussionmentioning

confidence: 99%

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

Adachi

Nakata

Tanabe

et al. 2021

IJMS

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“…Factors known to lead to false positives include plasma formulations produced by passing through cellulose membranes (albumin, γ globulin formulations), dialysis using cellulose membranes, and contamination by BDG in gauze [ 10 11 12 ]. BDG is widely distributed in the cell walls of fungi such as yeast, mushrooms, and mold, as well as in algae and higher plants.…”

Section: Discussionmentioning

confidence: 99%

“…The plasma portion of LRP was diluted to x (1-Ht) + x mL because the same amount of HES was added. Therefore, as a correction formula for this dilution, we multiplied the measured LRP value by (2-Ht) / (1-Ht) [ 4 11 ]. In other words, the upper limit of measurement using the PRP method is 900 pg / mL, and the lower limit of measurement is 0.6 pg / mL; however, the upper and lower limits can be increased by multiplying the measured value by the correction coefficient.…”

Section: Methodsmentioning

confidence: 99%

Basic Verification of β-D Glucan in Leukocyte-Rich Plasma for the Diagnosis of Deep Mycosis

et al. 2021

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Background Currently, supplementary serological testing for β-D glucan (BDG) is often selected to diagnose deep mycosis in care covered by the health insurance in Japan. The Wako method used by our center has low sensitivity, and different studies have used different cut-off values due to factors that cause false positives and false negatives. One possible cause of false negatives is the use of platelet-rich plasma (PRP) as the sample material. Because phagocytic white blood cells (WBC) are precipitated by centrifugation and only plasma is measured, it seems unlikely that the actual amount of BDG is being measured when using PRP. Further, a frequent cause of false positives is contamination from blood products and gauze containing BDG. To resolve these issues, the blood cell separator, hydroxyethyl starch, is used to precipitate only the red blood cells to obtain leukocyte-rich plasma (LRP). We hypothesized that it might be possible to improve the diagnostic rate of deep mycosis by measuring the BDG content of plasma containing WBC and fungal components and by comparing the BDG content of PRP and LRP measured simultaneously. Materials and Methods Healthy human blood, albumin-added blood, wrung-out gauze fluid-added blood, and fungal solution-added blood were prepared, and PRP and LRP were prepared using hydroxyethyl starch. The BDG content of each sample was measured using the Wako method and compared. In addition, PRP and LRP of fungal-added blood were Gram-stained and examined under a microscope, and the number of WBCs and phagocytosed fungi was counted visually and compared. Results Measuring the BDG content of LRP confirmed that there were no false positives with LRP, and in vitro experiments comparing albumin-added false-positive blood to fungal-added blood showed significant differences between PRP and LRP only in the fungal-added blood. Conclusion Calculating the BDG-ratio (LRP/PRP) by measuring both LRP and PRP may eliminate false positives and false negatives of true deep mycosis and improve the diagnostic rate.

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“…Schizophyllan (SPG) and lentinan (LNT), which are fungal BGs used as main ingredients in some medicinal preparations, will naturally give LAL-assay-positive results [ 15 ]. Some antibacterial, antiviral, antifungal, or blood products have also been reported to contain LAL-assay-positive substances [ 16 , 17 , 18 ]. These medicines and medical devices are often used in patients who are at high risk of developing deep mycosis.…”

Section: Introductionmentioning

confidence: 99%

Possibility of Japanese Cedar Pollen Causing False Positives in the Deep Mycosis Test

Kanno

Kim

Yamanaka

et al. 2021

IJMS

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Because Japanese cedar pollen (JCP) contains beta-1,3-d-glucan (BG), there is concern that its lingering presence in the atmosphere, especially during its scattering period, may cause false positives in the factor-G-based Limulus amebocyte lysate (LAL) assay used to test for deep mycosis (i.e., G-test). Hence, we examined whether the LAL assay would react positively with substances contained in JCP by using the G-test to measure JCP particles and extracts. BG was purified from the JCP extract on a BG-specific affinity column, and the percentage extractability was measured using three different BG-specific quantitative methods. The G-test detected 0.4 pg BG in a single JCP particle and 10 fg from a single particle in the extract. The percentage extractability of JCP-derived BG was not significantly different among the three quantitative methods. As the JCP particles should technically have been removed during serum separation, they should be less likely to be a direct false-positive factor. However, given that the LAL-assay-positive substances in the JCP extract were not distinguishable by the three BG-specific quantitative methods, we conclude that they may cause the background to rise. Therefore, in Japan false positives arising from JCP contamination should be considered when testing patients for deep mycosis.

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The Effect of Intravenous Gamma-globulin Reagents on the Measurement Results of (1→3)-β-D-glucan

Cited by 3 publications

References 5 publications

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

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

Basic Verification of β-D Glucan in Leukocyte-Rich Plasma for the Diagnosis of Deep Mycosis

Possibility of Japanese Cedar Pollen Causing False Positives in the Deep Mycosis Test

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