Faculty of Veterinary Medicine, Ludwig-Maximilians-University Munich, Schoenleutnerstr. 8, 85764, Oberschleissheim, Germany.
Institute for Infectious Diseases and Zoonoses, Faculty of Veterinary Medicine, Ludwig-Maximilians-University Munich, Veterinaerstraße 13, 80539, Munich, Germany.
Anal Bioanal Chem. 2021 Aug;413(19):4801-4813. doi: 10.1007/s00216-021-03436-y. Epub 2021 Jun 15.
Cytotoxic macrocyclic trichothecenes such as satratoxins are produced by chemotype S strains of Stachybotrys chartarum. Diseases such as stachybotryotoxicosis in animals and the sick building syndrome as a multifactorial disease complex in humans have been associated with this mold and its toxins. Less toxic non-chemotype S strains of S. chartarum are morphologically indistinguishable from chemotype S strains, which results in uncertainties in hazard characterization of isolates. To selectively identify macrocyclic trichothecene producing S. chartarum isolates, a set of sat14 gene-specific primers was designed and applied in a loop-mediated isothermal amplification (LAMP) assay using neutral red for visual signal detection. The assay was highly specific for S. chartarum strains of the macrocyclic trichothecene producing chemotype and showed no cross-reaction with non-macrocyclic trichothecene producing S. chartarum strains or 152 strains of 131 other fungal species. The assay's detection limit was 0.635 pg/rxn (picogram per reaction) with a reaction time of 60 min. Its high specificity and sensitivity as well as the cost-saving properties make the new assay an interesting and powerful diagnostic tool for easy and rapid testing.
细胞毒性大环多醚类如 satratoxins 是由链格孢菌化学型 S 菌株产生的。这种霉菌及其毒素与动物的 stachybotryotoxicosis 疾病和作为多因素疾病综合征的病态建筑综合征有关。形态上与化学型 S 菌株无法区分的毒性较低的非化学型 S 链格孢菌株，导致对分离物危害特征的不确定性。为了选择性地鉴定产生大环多醚类的链格孢菌分离物，设计了一组 sat14 基因特异性引物，并应用于环介导等温扩增（LAMP）检测，使用中性红进行可视信号检测。该检测法对产生大环多醚类的化学型链格孢菌菌株具有高度特异性，与不产生大环多醚类的链格孢菌菌株或 131 种其他 152 种真菌物种没有交叉反应。检测限为 0.635 pg/rxn（皮克/反应），反应时间为 60 分钟。该新检测法具有高特异性和敏感性以及节省成本的特点，使其成为一种有趣且强大的诊断工具，可用于简便快速的测试。
