Department of Pharmaceutical Sciences, Faculty of Pharmacy, The University of Jordan, Amman-Jordan, P.O. Box 11942, Amman, Jordan.
Department of Chemistry, Alfaisal University, Al Zahrawi Street, Al Maather, Al Takhassusi Road, Riyadh, 11533, Saudi Arabia.
Mikrochim Acta. 2019 Mar 8;186(4):230. doi: 10.1007/s00604-019-3313-0.
A colorimetric assay is presented for the detection of Stachybotrys chartarum proteases as biomarkers. The assay comprises a gold film acting as solid support and carrying an immobilized peptide substrate that is specific for S. chartarum protease. The substrate was conjugated to black magnetic nanoparticles (MNPs) to form a monolayer on the gold film. Hence, detection nanoprobe is black. If, however, the peptide-MNP fragments are cleaved by S. chartarum proteases present in a sample, the golden color of the detecting nanoprobe becomes apparent so that positive visual readout is enabled. The method was applied to the determination of S. chartarum in (spiked) environmental samples. The limit of detection ranges from 10 to 100 spores·mL depending on the kind of sample (culture, dust, mold and soil). Assay specificity was examined for Aspergillus flavus, Fusarium solani. Penicillin chrysogenum, and Saccharomyces cerevisiae, and negative readouts were observed visually for all samples, except for those also containing S. chartarum. Graphical abstract Schematic presentation of S. chartarum colorimetric nanoprobe.
本文提出了一种比色法检测拟青霉蛋白酶作为生物标志物的方法。该测定法包括作为固体支撑物的金膜和携带针对拟青霉蛋白酶的固定化肽底物。该底物与黑色磁性纳米颗粒(MNPs)缀合,在金膜上形成单层。因此,检测探针为黑色。然而,如果肽-MNP 片段被样品中存在的拟青霉蛋白酶切割,则检测探针的金色变得明显,从而实现阳性目视读数。该方法已应用于(污染)环境样品中拟青霉的测定。检测限范围为 10 到 100 个孢子·毫升,具体取决于样品类型(培养物、灰尘、霉菌和土壤)。对黄曲霉、茄病镰刀菌、青霉素和酿酒酵母进行了检测特异性检查,除了那些也含有拟青霉的样品外,所有样品均观察到阴性目视读数。