Laboratory for Immunopharmacology of Microbial Products, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi Hachioji, Tokyo 192-0392, Japan.
Department of Host Defense and Responses, Kagawa Nutrition University, 3-9-21 Chiyoda, Sakado, Saitama 350-0288, Japan.
Int J Mol Sci. 2021 Feb 21;22(4):2135. doi: 10.3390/ijms22042135.
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.
由于日本扁柏花粉(JCP)中含有β-1,3-d-葡聚糖(BG),人们担心其在空气中的持续存在,特别是在其散布期间,可能会导致基于因子 G 的鲎变形细胞溶解物(LAL)检测法(即 G 试验)对深部真菌感染的假阳性。因此,我们通过使用 G 试验来测量 JCP 颗粒和提取物,来检查 LAL 检测法是否会与 JCP 中所含的物质产生阳性反应。BG 是从 JCP 提取物中通过 BG 特异性亲和柱纯化的,使用三种不同的 BG 特异性定量方法测量其提取率。G 试验检测到单个 JCP 颗粒中有 0.4 pg 的 BG,从提取物中的单个颗粒中可检测到 10 fg 的 BG。JCP 来源的 BG 的提取率在三种定量方法之间没有显著差异。由于在血清分离过程中技术上应该已经去除了 JCP 颗粒,因此它们不太可能成为直接的假阳性因素。然而,鉴于 JCP 提取物中的 LAL 检测阳性物质不能通过三种 BG 特异性定量方法区分,我们推断它们可能会导致背景升高。因此,在日本,当检测患者是否患有深部真菌感染时,应考虑 JCP 污染引起的假阳性。