Galderma R&D, Sophia-Antipolis, France.
Department of Dermatology, Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland.
Mycoses. 2019 Jan;62(1):32-40. doi: 10.1111/myc.12843. Epub 2018 Oct 14.
Onychomycosis is the most prevalent nail disease and is mainly caused by two dermatophyte species Trichophyton rubrum and Trichophyton interdigitale with a frequency in the range of 80% and 20%, respectively. The secreted protease Sub6 of the subtilisin family, which was never detected in vitro growth conditions, was found to be a robust marker of onychomycosis.
The aim of this work was to detect tinea unguium using anti-Sub6 monoclonal antibodies in proteins extracted from clinical nail samples.
We produced monoclonal antibodies in mice using recombinant Sub6 as an antigen. Selected monoclonal antibodies were tested by Western blot analysis and ELISA on protein extracts from onychomycosis samples.
Several monoclonal antibodies used to quantify Sub6 in proteins extracted from clinical nail samples were produced and characterised. We showed that these antibodies were very specific and allowed the detection of T. rubrum and T. interdigitale in onychomycosis. Sub6 was detected in clinical samples infected by T. rubrum and not detected in nails with trauma and other diseases.
Anti-Sub6 monoclonal antibodies could be useful for a rapid diagnosis of tinea unguium and/or therapeutic survey of dermatophyte in onychomycosis by ELISA or an immunochromatography device such as a strip test.
甲真菌病是最常见的指甲疾病，主要由两种亲角质真菌引起，即红色毛癣菌和须癣毛癣菌，分别占 80%和 20%。从未在体外生长条件下检测到的枯草杆菌蛋白酶家族分泌的蛋白酶 Sub6 被发现是甲真菌病的一个强有力的标志物。
本研究旨在使用从临床指甲样本中提取的蛋白质中的抗 Sub6 单克隆抗体来检测甲癣。
我们使用重组 Sub6 作为抗原在小鼠中产生单克隆抗体。选择的单克隆抗体通过 Western blot 分析和 ELISA 在甲真菌病样本的蛋白质提取物上进行测试。
制备并鉴定了用于定量从临床指甲样本中提取的蛋白质中的 Sub6 的几种单克隆抗体。我们表明，这些抗体非常特异，可检测甲真菌病中的红色毛癣菌和须癣毛癣菌。Sub6 可在红色毛癣菌感染的临床样本中检测到，而在创伤和其他疾病的指甲中未检测到。
抗 Sub6 单克隆抗体可通过 ELISA 或免疫层析设备（如条带测试）用于甲真菌病的快速诊断和/或对亲角质真菌的治疗监测。
